Absorbance Microplate Reader

ELx808™
Operator’s Manual
 
ELx808™

Absorbance Microplate Reader, All Models

Operator’s Manual

April 2013
© 2013
Part Number 7341000
Revision P
BioTek® Instruments, Inc.
ii | Preface

Notices
BioTek® Instruments, Inc.
Highland Park, P.O. Box 998
Winooski, Vermont 05404-0998 USA

All Rights Reserved

© 2013, BioTek® Instruments, Incorporated. No part of this publication may be

reproduced, transcribed, or transmitted in any form, or by any means electronic or
mechanical, including photocopying and recording, for any purpose other than the
purchaser’s use without written permission of BioTek Instruments, Inc.

Trademarks

BioTek® is a registered trademark, and ELx808™, 4-Zone™ and Gen5™ are trademarks
of BioTek Instruments, Inc. Microsoft®, Windows®, and Excel® are either registered
trademarks or trademarks of Microsoft Corporation in the United States and/or other
countries.
All other trademarks are the property of their respective holders.

Restrictions and Liabilities

Information in this document is subject to change and does not represent a

commitment by BioTek Instruments, Inc. Changes made to the information in this
document will be incorporated in new editions of the publication. No responsibility is
assumed by BioTek for the use or reliability of software or equipment that is not
supplied by BioTek or its affiliated dealers.

BioTek Instruments, Inc.

 Contents | iii

Contents
Notices ........................................................................................... ii 
All Rights Reserved ...................................................................... ii 
Trademarks ................................................................................ ii 
Restrictions and Liabilities ............................................................ ii 
Contents ........................................................................................ iii 
Contact Information ...................................................................... viii 
Customer Service and Sales....................................................... viii 
Service/TAC ............................................................................ viii 
European Coordination Center/Authorized European Representative viii 
Revision History .............................................................................. ix 
Document Conventions ................................................................... xii 
Intended Use Statement ................................................................. xii 
Quality Control .............................................................................. xii 
Warranty & Product Registration...................................................... xiii 
Repackaging and Shipping .............................................................. xiii 
Warnings ..................................................................................... xiii 
Hazards ....................................................................................... xiv 
Precautions ................................................................................... xv 
CE Mark ....................................................................................... xvi 
Directive 2004/108/EC: Electromagnetic Compatibility................... xvi 
Directive 2006/95/EC Low Voltage (Safety) .................................. xvi 
Directive 2002/96/EC: Waste Electrical and Electronic Equipment... xvii 
Directive 98/79/EC: In Vitro Diagnostics (if labeled for this use) .... xvii 
Electromagnetic Interference and Susceptibility ................................ xvii 
USA FCC CLASS A ................................................................... xvii 
Canadian Department of Communications Class A ........................ xvii 
User Safety ................................................................................. xviii 
Safety Symbols ............................................................................. xix 
Chapter 1: Introduction.......................................................................1
Introducing the ELx808 Absorbance Microplate Reader ......................... 2
Reader Variations ....................................................................... 2
Hardware Features .......................................................................... 2
Software Features ........................................................................... 3
Package Contents ............................................................................ 3
Optional Accessories ........................................................................ 3
Specifications ................................................................................. 4
Microplates ................................................................................ 4
Electrical ................................................................................... 4
Physical .................................................................................... 4
Environmental ........................................................................... 5
Hardware .................................................................................. 5
Reading Speeds ......................................................................... 5

ELx808 Operator’s Manual

iv | Preface

Standard Model: ELx808 ............................................................. 5

Ultraviolet/Incubator Model – ELx808IU ......................................... 6
Product Support & Service ................................................................ 7
Contacting the Technical Assistance Center .................................... 7
Returning Instruments for Service/Repair ...................................... 7
Chapter 2: Installation ........................................................................9
Product Registration ....................................................................... 10
1: Unpack and Inspect the Instrument .............................................. 10
2: Select the Operating Environment ................................................ 11
3: Install the Filter Wheel ................................................................ 12
4: Check/Adjust the Power Input Voltage Setting ................................ 14
5: Connect Power ........................................................................... 14
6: (Optional) Connect Printer ........................................................... 15
Printers.................................................................................... 16
7: Turn on Reader and Run System Test............................................ 16
8: Check/Adjust the Reader’s Filter Table .......................................... 17
9: Configure Reader Settings ........................................................... 18
SETUP Options .......................................................................... 18
OUTPUT Options........................................................................ 19
REPORT Type............................................................................ 19
READ Options ........................................................................... 20
Read Speed .............................................................................. 21
10: Install Software/Connect to a Computer (Optional) ........................ 21
Attach the Cable ....................................................................... 21
Install Gen5 Software on the Host Computer ................................. 21
Establish Communication ............................................................ 22
Changing the Baud Rate on the ELx808 ........................................ 22
11: Verify Performance ................................................................... 23
Before Repackaging the Instrument .................................................. 24
Chapter 3: Operation ......................................................................... 25
Getting Started with Gen5 ............................................................... 27
Introduction .................................................................................. 28
The Keypad .............................................................................. 29
The Startup Screen.................................................................... 30
The Main Menu Screen ............................................................... 30
Quick Read ............................................................................... 31
Defining Assays ............................................................................. 32
Selecting an Assay .................................................................... 32
Assay Name ............................................................................. 32
Defining the Method, Map, Formula, and Curve.............................. 33
METHOD .................................................................................. 34
Read Type ........................................................................... 34
Delay in First Read ............................................................... 34
Incubation Temperature ........................................................ 35
Single or Dual Wavelength ..................................................... 35

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 Contents | v

MEAS Selection .................................................................... 36

Number of Kinetic Reads/Kinetic Duration Selection .................. 36
Kinetic Interval ................................................................... 36
Kinetic Number of Reads ....................................................... 37
Kinetic Duration ................................................................... 37
Shake Mode Selection ........................................................... 37
Shake Time ......................................................................... 38
Shake Speed ....................................................................... 38
Kinetic Data Analysis Selection ............................................... 38
Number of Kinetic Points Selection .......................................... 39
Onset OD Selection............................................................... 39
Linear Scanning ................................................................... 39
MAP ........................................................................................ 40
Map Generation.................................................................... 41
Mapping Direction ................................................................ 42
Replication Direction ............................................................. 42
Examples of Mapping & Replication Directions .......................... 43
Start Mapping at Well Location ............................................... 44
Selecting a Blank Map ........................................................... 44
Blank Map Definitions............................................................ 45
Constant Blank Value Entry.................................................... 46
Number of Blanks ................................................................. 46
Selecting a Blank Location ..................................................... 46
Number of Standards ............................................................ 47
Number of Standard Replicates .............................................. 47
Average Standards ............................................................... 47
Standard Concentration ......................................................... 48
Reuse of Standard Curves ..................................................... 49
Number of Controls .............................................................. 50
Control Type ........................................................................ 50
Number of Control Replicates ................................................. 51
Location of Controls .............................................................. 51
Valid Well Locations .............................................................. 51
Number of Samples .............................................................. 52
Number of Sample Replicates ................................................. 52
Sample Location................................................................... 52
FORMULA ................................................................................. 53
Validation Formula Examples ................................................. 53
Formula Type ...................................................................... 53
Validation Type Selection ....................................................... 55
Formula Entry ...................................................................... 55
Number of Required Controls/Blanks ....................................... 57
Cutoff Formulas ................................................................... 58
Greyzone Entry .................................................................... 59
Positive/Negative Calls for Cutoff ............................................ 59

ELx808 Operator’s Manual

vi | Preface

Examples ............................................................................ 60
Transformation Formulas ....................................................... 61
Transformation Formula Definition .......................................... 61
Transformation Scope Variable ............................................... 62
Another Transformation Example ............................................ 63
CURVE ..................................................................................... 64
Curve Fit ............................................................................. 64
Edit Standard Outliers ........................................................... 65
Axis Selection ...................................................................... 66
Extrapolation of Unknowns .................................................... 66
Panel Assays ............................................................................ 67
Reading a Microplate ...................................................................... 70
Select Assay ............................................................................. 70
Run-Time Prompts..................................................................... 70
Enter Number of Samples ........................................................... 71
Enter Plate ID ........................................................................... 71
Enter Sample ID ....................................................................... 71
Prompts for Well Location ........................................................... 72
Beginning the Plate Read ............................................................ 72
Printing Reports and Assay Lists ....................................................... 73
Result ...................................................................................... 74
Editing Standard Outliers ............................................................ 74
Printing Results ......................................................................... 75
Map......................................................................................... 75
Assay ...................................................................................... 76
List ......................................................................................... 76
Recommendations for Optimum Performance ..................................... 76
Chapter 4: Instrument Qualification ..................................................79
Overview ...................................................................................... 80
Recommended Qualification Schedule ............................................... 80
Test Descriptions ........................................................................... 81
System Test ............................................................................. 81
Incubator Self Test ............................................................... 84
Checksum Test ......................................................................... 84
Absorbance Plate Test ................................................................ 85
Empty Carrier Test .................................................................... 88
Liquid Tests .............................................................................. 88
Perform the Tests........................................................................... 89
System Test ............................................................................. 89
Checksum Test ......................................................................... 89
Absorbance Plate Test ................................................................ 89
Define the Test Plate Parameters ........................................... 89
Run the Plate Test ............................................................... 90
Empty Carrier Test .............................................................. 91
Liquid Tests .............................................................................. 92
Prepare Test Solutions ......................................................... 92

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 Contents | vii

Stock Solution Formulation ......................................................... 92

Solution A ........................................................................... 92
Solution B ........................................................................... 93
Perform Liquid Test 1 ................................................................. 94
Calculations ......................................................................... 94
Accuracy Specifications ......................................................... 95
Perform Liquid Test 2 ................................................................. 95
Perform the Linearity Test (Test A) ......................................... 96
Calculate Repeatability (Test B) .............................................. 97
Channel-to-Channel Variation and Alignment (Test C)................ 97
Perform Liquid Test 3 (“UV” Models Only) ..................................... 98
Perform the Test .................................................................. 99
Calculate Repeatability .......................................................... 99
Calculate Linearity .............................................................. 100
Chapter 5: Preventive Maintenance ................................................. 101
Recommended Maintenance Schedule ............................................. 102
Warnings and Precautions ............................................................. 102
Clean Exposed Surfaces ................................................................ 103
Inspect and Clean the Wavelength Filters ........................................ 104
(Optional) Lubricate Robotic Components ........................................ 105
Robotic Motor Adjustment Procedure .......................................... 107
Replace the Lamp and Clean the Contacts........................................ 107
Decontamination .......................................................................... 111
Purpose ................................................................................. 111
Tools and Supplies .................................................................. 112
Decontamination Procedure ...................................................... 112
Chapter 6: Troubleshooting and Error Codes ...................................115
Overview .................................................................................... 116
Diagnostics ............................................................................ 116
Terms Used in the Error Codes Tables ........................................ 116
Error Codes (General) ................................................................... 117
Error Codes (Fatal) ....................................................................... 121
Appendix A: Sample Reports ........................................................... 123
Appendix B: Instructions for Programming a New Assay ................131
Appendix C: Adjusting the Power Input Voltage Setting ................. 141

ELx808 Operator’s Manual

viii | Preface

Contact Information

™ See also Product Support and Service on page 7.

BioTek® Instruments, Inc.

Highland Park, P.O. Box 998
Winooski, Vermont 05404-0998 USA

Customer Service and Sales

Internet: www.biotek.com
Phone: 888-451-5171 (toll free in the U.S.)
802-655-4740 (outside the U.S.)
Fax: 802-655-7941
E-Mail: customercare@biotek.com

Service/TAC

Phone: 800-242-4685 (toll-free in the U.S.)

802-655-4740 (outside the U.S.)
Fax: 802-654-0638
E-Mail: tac@biotek.com

European Coordination Center/Authorized European

Representative

BioTek® Instruments GmbH

Kocherwaldstrasse 34
D-74177 Bad Friedrichshall
Germany
Internet: www.biotek.de
Phone: +49 (0) 7136 9680
Fax: +49 (0) 7136 968 111
E-Mail: info@biotek.de

BioTek Instruments, Inc.

 Revision History | ix

Revision History

Rev Date Changes

A 1/96 First Release
B 5/96 Revised reader specifications. Added Scanning method to software.
C 1/97 Added Panel and Reuse of Standard Curve, and revised serial port pin-out
description.
D 2/98 Changed Chapter 4, Performance Verification to reflect changes to the Universal
Test Plate. Added Liquid Tests 1 and 2. Changed Appendix C: Error Codes.
E 8/98 Added printer information. Updated Appendix B: Computer Control.
F 3/99 Changed European address. Corrected printer compatibility. Corrected the
sequence of steps for processing a curve-fit method.
G 7/99 Added comment about blanking, P-Down and P-Across being inactive in this
version of software. Clarified the need for at least one sample to be defined on a
plate. Added scanning computer control commands.
H 5/00 Updated Chapter 4- Performance Verification to include a Liquid Test 3 to verify
340 nm instruments. Corrected the voltage range for Range 2 in the
Introduction. Corrected Incubation Temperature Control range to 6 deg above
ambient. Changed Note on page 4-7 dealing with air readings during Self-Test.
Clarified Liquid Test 1. Changed Table 4-1 to include additional maintenance
items. Reworked Liquid Test 2.
I 7/02 Updated contact information (pages iii, 1-8, 1-9, 2-8, 2-20, and 4-13). Added
IQ/OQ/PQ procedures and updated liquid testing information (Chapter 4).
Revised lamp replacement procedure (page 2-15).
J 11/03 Preface: Updated contact information in Notices (page iii). Added Document
Conventions (page vi). Updated Warnings section (pages vii and viii). Updated
Electromagnetic Compatibility section (pages ix and x). Added the following
safety symbols and text: “Consult instructions for use” (page xii) “In vitro
diagnostic medical device” (page xii) “Separate collection for (disposal of)
electrical and electronic equipment” (page xiii). Expanded the Intended Use
Statement (page xiv).
Chapter 1: Updated contact information in Technical Support. Removed About
This Manual section (page 1-5). Added “Absorbance Test Plate” to the Optional
Accessories list (page 1-8). Clarified lamp replacement procedure (Chapter 2,
pages 2-14 to 2-16). Clarified description of cutoff formulas (Chapter 3, pages 3-
42 to 3-44).
Chapter 4: Changed title to “Performance Verification and IQ, PQ, OQ Tests.”
Added IQ/PQ/OQ test procedure information. Clarified procedures for liquid
tests.
J 11/03 Revised decontamination instructions and added cleaning procedure (Appendix
A). Added KC4 startup information to Appendix B. Added new Appendix E with
two examples of assay kit instructions and directions for programming an assay.
General: Edited and formatted text. Modified appearance of display screens.
Standardized the presentation of significant digits. Changed “Abs” to “OD”
throughout.

ELx808 Operator’s Manual

x | Preface

Rev Date Changes

K 4/05 Updated the cover with a current photo of the instrument. Updated contact
information and warranty. Changed “Automated Microplate Reader” to
“Absorbance Microplate Reader” throughout. Changed “Universal Test Plate” to
“Absorbance Test Plate” throughout. Removed references to internal barcode
scanner and “R” models. Removed references to General Formulas with respect
to open assays. Updated specifications to match product spec rev D. Added text
from rev J1 insert (Fuse Installation/Replacement). Updated the installation
instructions in Chapter 2 to better reflect actual practice. Updated the
IQ/OQ/PQ/PM steps in Chapter 4 to better reflect actual practice. Created
Appendix F to provide instructions for adjusting the line input voltage range and
replacing fuses for instruments with an older-style power input module than the
one described in Chapter 2.
L 4/06 Updated the manual primarily to support the introduction of Gen5™ software. In
general, changed ‘Bio-Tek’ to ‘BioTek,’ and added Gen5 instructions wherever
KCjunior™ and KC4™ instructions were present.
Cover: Updated BioTek logo to new graphic.
Preface: Added Contact Information section, updated Hazards, Precautions, and
safety standards. Removed Registration Card and Warranty (these items ship
separately).
Chapter 1: Added external power supply to Hardware Features, Package
Contents, and Specifications. Also added ‘Reading Speeds’ section to specs.
Replaced Technical Support section with one-page Product Support & Service
section.
Chapter 2: Added instructions for connecting the external power supply in
Connect Power section. Moved instructions in Check/Adjust the Power Input
Voltage Setting section to Appendix F.
Chapter 4: Changed chapter title from ‘Performance Verification, Periodic
Maintenance, and IQ, PQ, OQ Tests’ to ‘Instrument Verification’. Moved
maintenance instructions to new Chapter 5, Preventive Maintenance (see below).
For Liquid Tests 1, 2, and 3, added recommendation to shake the plate for four
minutes (or wait for 20 minutes) after pipetting the diluted test solution, before
reading the plate.
Chapter 5 (new chapter): In Recommended Maintenance table, removed 3-
month interval for cleaning the lamp contacts and changed ‘Replace the Lamp’ to
‘Replace the Lamp and Clean the Contacts.’ Added cleaning instructions from
Appendix A, Decontamination.
Appendix B: Added new section: ‘Controlling the Reader with Gen5’.
M 3/08 General: Applied the BioTek template to the manual. Changed “Bio-Tek” to
“BIOTEK” in startup screens to reflect this change in the reader’s basecode
software.
Preface: Added Microsoft®, Windows®, Windows XP, Windows 2000, Windows
Vista™, and Excel® to the “Trademarks” section. In the “Contacting BioTek
Instruments” section, updated the TAC fax number and added “Authorized
European Representative” to the “European Coordination Center” heading.
Chapter 1: Added part number disclaimer to “Package Contents” and “Optional
Accessories” and updated the TAC fax number in the “Product Support and
Service” section.
Chapter 2: Added a packaging disclaimer to the section “Unpack and Inspect the
Instrument.”
Chapter 3: Enhanced section on the keypad with graphics that illustrate
components of the ELx808™keypad/ display. Updated “Panel Assays” section. In

BioTek Instruments, Inc.

 Revision History | xi

Rev Date Changes

the “Printing Reports and Assay Lists” section, added a note containing an
explanation of the OUT of range indication on reports.
Chapter 4: Clarified the description of the System Test Report to state that the
incubation test is done ‘separately’ and that the overall system test results given
at the bottom of the test report refer only to the Optics portion of the test.
Capitalized the “l” in “μl” and “ml.”
Chapter 5: Merged this chapter with Appendix A, Decontamination.
Reformatted and renamed Appendix C, Error Codes, as “Chapter 6, Error Codes”.
Added a “Fatal Errors” section, and added information on Error Codes 2311 to
2386.
Renamed Appendix B, Computer Control, as Appendix A.
Renamed Appendix D, Reports, as Appendix B, Sample Reports. In the
“Overview,” added a note containing an explanation of the OUT of range
indication on reports.
Renamed Appendix E, Programming a New Assay as Appendix C.
Renamed Appendix F, Adjusting the Power Input Voltage Setting as Appendix D.
N 6/11 Throughout: Removed references to outdated software KC4 and KCjunior.
Preface: Updated Trademark. Updated Warnings and Precautions. Updated
Intended Use statement.
Chapter 2: Removed Parallel and Serial Port Pinout charts. Added Install
Software/Connect to Computer.
Chapter 3: Added “Getting Started with Gen5”. Gave Baud Rate information for
Gen5. Removed references to outdated software Extensions.
Chapter 4: Removed Installation Qualification. Updated Autocal Analysis.
Updated sample Absorbance Test Plate Results. Updated Stock Solution
Formulation for Liquid Tests 1 and 2. Removed outdated instructions for creating
former Solution A: 10x Concentrate PBS from Liquid Test 3. Simplified test
procedure as a result.
Chapter 5: Updates lamp replacement instructions (added text from former
revision M1 insert).
Chapter 6: Added error code 1201.
Appendices: Removed former Appendix A: Computer Control.
O 9/12 General: Updated Gen5 instructions to reflect terminology in Gen5 v2.x.
Preface: Updated the Intended Use Statement and the heading for the In
Vitro Diagnostics directive to refer to the instrument’s IVD label (if one
exists). Added ‘Service’ and ‘Accessories’ hazard warnings. Added ‘Spare
Parts’ precaution.
Chapter 1, Introduction: Updated part number for the power supply.
Chapter 2, Installation: Added note that improper packaging that results in
damage to the instrument may lead to additional charges.
Chapter 4, Qualification: Added a note to the Absorbance Test Plate section
stating that the test tests the accuracy and repeatability of the reader to 2.500
OD. It does not indicate a PASS or FAIL above 2.500 OD.
P 4/13 Preface: Added EN 61010-2-081 and EN 61010-2-010 to Directive 2006/95/EC
Low Voltage (Safety) and EN 61010-2-101 to Directive 98/79/EC In Vitro
Diagnostics.

ELx808 Operator’s Manual

xii | Preface

Document Conventions
See also Safety Symbols on page xix.

This icon calls attention to important safety notes.

A Warning indicates the potential for bodily harm and

Warning!
tells you how to avoid the problem.

A Caution indicates potential damage to the instrument

Caution
and tells you how to avoid the problem.

Note: Bold text is primarily used for emphasis.

This icon calls attention to important information.

Intended Use Statement

• The ELx808 is an absorbance microplate reader. The performance characteristics of the

data reduction software have not been established with any laboratory diagnostic
assay. The user must evaluate this instrument and (if used) PC-based software in
conjunction with their specific assay(s). This evaluation must include the confirmation
that performance characteristics for the specific assay(s) are met.
• If the instrument has an “IVD” label it may be used for clinical and non-clinical
purposes, including research and development. If there is no such label the instrument
may only be used for research and development or other non-clinical purposes.

Quality Control
It is considered good laboratory practice to run laboratory samples according to instruc-
tions and specific recommendations included in the assay package insert for the test to be
conducted. Failure to conduct Quality Control checks could result in erroneous test data.

BioTek Instruments, Inc.

 Warranty & Product Registration | xiii

Warranty & Product Registration

Take a moment to review the warranty information that shipped with your product.
Please also register your product with BioTek to ensure that you receive important
information and updates about the product(s) you have purchased. You can register online
through the Customer Resource Center (CRC) at www.biotek.com or by calling 888-451-
5171 or 802-655-4740.

Repackaging and Shipping

If you need to ship the instrument to BioTek for service or repair,

contact BioTek for a Return Materials Authorization (RMA)
number, and be sure to use the original packing materials. Other
forms of commercially available packaging are not recommended
and can void the warranty. If the original packing materials have
been damaged or lost, contact BioTek for replacement packing.

Warnings

Operate the instrument on a level, stable surface away from excessive humidity.
Bright light or strong incandescent light can reduce the linear performance range
of the instrument.
Measurement values may be affected by extraneous particles (such as dust) in the
microplate wells. A clean work area is necessary to ensure accurate readings.
When operated in a safe environment according to the instructions in this
document, there are no known hazards associated with the instrument. However,
the operator should be aware of certain situations that could result in serious
injury; these may vary depending on the instrument model. See Hazards and
Precautions.

ELx808 Operator’s Manual

xiv | Preface

Hazards
The following hazard warnings are provided to help avoid injury:

Warning! Internal Voltage. Always turn off the power switch and unplug the
power cord or power supply before cleaning the outer surface of the instrument or
removing its top case.
Warning! Power Rating. The instrument’s power supply or power cord must be
connected to a power receptacle that provides voltage and current within the
specified rating for the system. Use of an incompatible power receptacle may
produce electrical shock and fire hazards.
Warning! Electrical Grounding. Never use a plug adapter to connect primary
power to the external power supply. Use of an adapter disconnects the utility
ground, creating a severe shock hazard. Always connect the power cord directly to
an appropriate receptacle with a functional ground.
Warning! Service. Only qualified technical personnel should perform service
procedures on internal components.
Warning! Accessories. Only accessories that meet the manufacturer’s
specifications shall be used with the instrument.
Warning! Liquids. Avoid spilling liquids on the reader; fluid seepage into
internal components creates a potential for shock hazard or instrument damage.
Wipe up all spills immediately. Do not operate the instrument if internal
components have been exposed to fluid.
Warning! Unspecified Use. Failure to operate this equipment according to the
guidelines and safeguards specified in this manual could result in a hazardous
condition.
Warning! Software Quality Control. The operator must follow the
manufacturer’s assay package insert when modifying software parameters and
establishing reading methods. Failure to conduct quality control checks could
result in erroneous test data.
Warning! Reader Data Reduction Protocol. When the reader is operated in
standalone mode the onboard assay software will flag properly defined controls
when they are out of range. The software will present the data with the
appropriate error flags for the operator to determine control and assay validity.
When operated via computer control, no limits are applied to the raw
measurement data. All information exported via computer control must be
thoroughly analyzed by the operator.
Warning! Hot Surface. The lamp is hot when the instrument is turned on. Turn
off the reader and allow the lamp to cool for at least 15 minutes before attempting
to replace it.

BioTek Instruments, Inc.

 Precautions | xv

Warning! Potential Biohazards. Some assays or specimens may pose a

biohazard. Adequate safety precautions should be taken as outlined in the assay’s
package insert. Always wear safety glasses and appropriate protective equipment,
such as chemically resistant rubber gloves and apron.

Precautions
The following precautions are provided to help avoid damage to the instrument:

Caution: Service. The instrument should be serviced by BioTek-authorized

personnel. Only qualified technical personnel should perform service procedures
on internal components.
Caution: Spare Parts. Only approved spare parts should be used for
maintenance. The use of unapproved spare parts and accessories may result in a
loss of warranty and potentially impair instrument performance or cause damage
to the instrument.
Caution: Environmental Conditions. Do not expose the system to temperature
extremes. For proper operation, ambient temperatures should remain within the
range listed in the Specifications section. Performance may be adversely affected
if temperatures fluctuate above or below this range. Storage temperature limits are
broader.
Caution: Sodium Hypochlorite. Do not expose any part of the instrument to the
recommended diluted sodium hypochlorite solution (bleach) for more than 20
minutes. Prolonged contact may damage the instrument surfaces. Be certain to
rinse and thoroughly wipe all surfaces.
Caution: Power Supply. Use only the power supply shipped with the
instrument. Operate the power supply within the range of line voltages listed on it.
Caution: Warranty. Failure to follow preventive maintenance protocols may
void the warranty.
Caution: Disposal. This instrument contains printed circuit boards and wiring
with lead solder. Dispose of the instrument according to Directive 2002/96/EC,
“on waste electrical and electronic equipment (WEEE)” or local ordinances.
Caution: Electromagnetic Environment. Per IEC 61326-2-6 it is the user’s
responsibility to ensure that a compatible electromagnetic environment for this
instrument is provided and maintained in order that the device will perform as
intended.
Caution: Electromagnetic Compatibility. Do not use this device in close
proximity to sources of strong electromagnetic radiation (e.g., unshielded
intentional RF sources), because these may interfere with the proper operation.

ELx808 Operator’s Manual

xvi | Preface

CE Mark

Based on the testing described below and information

contained herein, this instrument bears the CE mark.

™ See the Declaration of Conformity for more information.

Directive 2004/108/EC: Electromagnetic Compatibility

Emissions—Class A

The system has been type-tested by an independent, accredited testing laboratory

and found to meet the requirements of EN 61326-1: Class A for Radiated Emissions
and Line Conducted Emissions. Verification of compliance was conducted to the
limits and methods of EN 55011 – CISPR 11, Class A. In a domestic environment it
may cause radio interference, in which case you may need to mitigate the
interference.

Immunity

The system has been type-tested by an independent, accredited testing laboratory

and found to meet the requirements of EN 61326-1 and EN 61326-2-6 for Immunity.
Verification of compliance was conducted to the limits and methods of the
following:
EN 61000-4-2 Electrostatic Discharge
EN 61000-4-3 Radiated EM Fields
EN 61000-4-4 Electrical Fast Transient/Burst
EN 61000-4-5 Surge Immunity
EN 61000-4-6 Conducted Disturbances from RFI
EN 61000-4-11 Voltage Dips, Short Interruptions and Variations

Directive 2006/95/EC Low Voltage (Safety)

The system has been type-tested by an independent testing laboratory and was found
to meet the requirements of this Directive. Verification of compliance was conducted to
the limits and methods of the following:
EN 61010-1, “Safety requirement for electrical equipment for measurement, control and
laboratory use. Part 1, General requirements.”
EN 61010-2-081, “Particular requirements for automatic and semi-automatic laboratory
equipment for analysis and other purposes.”
EN 61010-2-010, “Particular requirements for laboratory equipment for the heating of
materials.”

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 Electromagnetic Interference and Susceptibility | xvii

Directive 2002/96/EC: Waste Electrical and Electronic Equipment

Disposal Notice: This instrument contains printed circuit boards and wiring with
lead solder. Dispose of the instrument according to Directive 2002/96/EC, “on
waste electrical and electronic equipment (WEEE)” or local ordinances.

Directive 98/79/EC: In Vitro Diagnostics (if labeled for this use)

• Product registration with competent authorities

• Traceability to the U.S. National Institute of Standards and Technology (NIST)
• EN 61010-2-101, “Particular requirements for in vitro diagnostic (IVD) medical
equipment.”

Electromagnetic Interference and Susceptibility

USA FCC CLASS A

RADIO AND TELEVISION INTERFERENCE

NOTE: This equipment has been tested and found to comply with the limits for a Class
A digital device, pursuant to Part 15 of the FCC Rules. These limits are designed to
provide reasonable protection against harmful interference when the equipment is
operated in a commercial environment. Like all similar equipment, this equipment
generates, uses, and can radiate radio frequency energy and, if not installed and used
in accordance with the instruction manual, may cause harmful interference to radio
communications. Operation of this equipment in a residential area is likely to cause
interference, in which case the user will be required to correct the interference at their
own expense.
In order to maintain compliance with FCC regulations, shielded cables must be used
with this equipment. Operation with non-approved equipment or unshielded cables is
likely to result in interference to radio and television reception.

Canadian Department of Communications Class A

This digital apparatus does not exceed Class A limits for radio emissions from digital
apparatus set out in the Radio Interference Regulations of the Canadian Department of
Communications.
Le present appareil numerique n'émet pas du bruits radioelectriques depassant les
limites applicables aux appareils numerique de la Class A prescrites dans le Reglement
sur le brouillage radioelectrique edicte par le ministere des Communications du
Canada.

ELx808 Operator’s Manual

xviii | Preface

User Safety
This device has been type-tested by an independent laboratory and found to meet the
requirements of the following:
• Underwriters Laboratories UL 61010-1, “Safety requirements for electrical
equipment for measurement, control and laboratory use; Part 1: General
requirements.”
• Canadian Standards Association CAN/CSA C22.2 No. 61010-1, “Safety
requirements for electrical equipment for measurement, control and laboratory
use; Part 1: General requirements.”
• EN 61010 Standards, see CE Mark starting on page xvi.

BioTek Instruments, Inc.

 Safety Symbols | xix

Safety Symbols
Some of these symbols may appear on the instrument or accessories:

Alternating current Both direct and alternating current

Courant alternatif Courant continu et courant alternatif
Wechselstrom Gleich - und Wechselstrom
Corriente alterna Corriente continua y corriente alterna
Corrente alternata Corrente continua e corrente alternata
Direct current Earth ground terminal
Courant continu Borne de terre
Gleichstrom Erde (Betriebserde)
Corriente continua Borne de tierra
Corrente continua Terra (di funzionamento)
On (Supply) Protective conductor terminal
Marche (alimentation) Borne de terre de protection
Ein (Verbindung mit dem Netz) Schutzleiteranschluss
Conectado Borne de tierra de protección
Chiuso Terra di protezione
Off (Supply) Caution (refer to accompanying documents)
Arrêt (alimentation) Attention (voir documents
Aus (Trennung vom Netz) d’accompanement)
Desconectado Achtung siehe Begleitpapiere
Aperto (sconnessione dalla Atención (vease los documentos incluidos)
rete di alimentazione) Attenzione, consultare la doc annessa
Warning, risk of electric shock Warning, risk of crushing or pinching
Attention, risque de choc Attention, risque d’écrasement et
électrique pincement
Gefährliche elektrische schlag Warnen, Gefahr des Zerquetschens und
Precaución, riesgo de sacudida Klemmen
eléctrica Precaución, riesgo del machacamiento y
Attenzione, rischio di scossa sejeción
elettrica Attenzione, rischio di schiacciare ed
intrappolarsi
Warning, hot surface Warning, potential biohazards
Attention, surface chaude Attention, risques biologiques potentiels
Warnen, heiße Oberfläche Warnung! Moegliche biologische Giftstoffe
Precaución, superficie caliente Atención, riesgos biológicos
Attenzione, superficie calda Attenzione, rischio biologico

ELx808 Operator’s Manual

xx | Preface

In vitro diagnostic medical Separate collection for electrical and

device electronic equipment
Dispositif médical de Les équipements électriques et
diagnostic in vitro électroniques font l’objet d’une collecte
Medizinisches In-Vitro- sélective
Diagnostikum Getrennte Sammlung von Elektro- und
Dispositivo médico de Elektronikgeräten
diagnóstico in vitro Recogida selectiva de aparatos eléctricos y
Dispositivo medico diagnostico electrónicos
in vitro Raccolta separata delle apparecchiature
elettriche ed elettroniche
Consult instructions for use Laser radiation: Do not stare into beam
Consulter la notice d’emploi Rayonnement laser: Ne pas regarder
Gebrauchsanweisung beachten dans le faisceau
Consultar las instrucciones de Laserstrahlung: Nicht in den strahl
uso blicken
Consultare le istruzioni per uso Radiación de láser: No mire fijamente al
rayo
Radiazione di laser: Non stare nel fascio

BioTek Instruments, Inc.

Chapter 1

Introduction

This chapter introduces the ELx808 Absorbance Microplate Reader

and describes its hardware and software features, and technical
specifications. Instructions for contacting BioTek are provided on
page 7.

ELx808 Absorbance Microplate Reader .................................. 2

Reader Variations .......................................................... 2
Hardware Features ............................................................. 2
Software Features .............................................................. 3
Package Contents .............................................................. 3
Optional Accessories........................................................... 3
Specifications .................................................................... 4
Microplates ................................................................... 4
Electrical ...................................................................... 4
Physical ....................................................................... 4
Environmental .............................................................. 5
Hardware ..................................................................... 5
Reading Speeds ............................................................ 5
Standard Model: ELx808 ................................................ 5
Incubator/Ultraviolet Model: ELx808IU ............................. 6
Product Support and Service ............................................... 7
Contacting the Technical Assistance Center ....................... 7
Returning Instruments for Service/Repair ......................... 7
2 | Chapter 1: Introduction

ELx808 Absorbance Microplate Reader

BioTek’s ELx808 is an eight-channel reader-assay system. The reader can serve as a
standalone system, or can be controlled via BioTek’s Gen5 software.
Designed to automatically perform endpoint and kinetic analysis, the reader can measure
the optical density of solutions in 96-well microplates between 380 nm and 900 nm. The
UV option measures down to 340 nm.
The reader features superior optical specifications, with an extended dynamic range of up
to 4.000 absorbance units.
The instrument’s onboard processor, 2-line x 24-character LCD screen, and membrane
keys allow easy definition and management of assay protocols, templates, formulas, and
data. Results can be output in a printed report format, or exported for use in a variety of
ELISA-based data manipulation applications.

Reader Variations

• ELx808: Absorbance Microplate Reader (standard model).

• ELx808IU: Reader with incubation and UV capability. The ELx808IU has a four-
zone incubation chamber that controls temperatures up to 50°C, and is capable of
reading plates between 340 nm and 900 nm wavelengths.

Hardware Features

• Eight optics channels, with an additional reference channel

• A wavelength range of 380-900 nm (ELx808), 340-900 nm (ELx808IU)
• A user-accessible, 6-position filter wheel
• A 2-line x 24-character LCD display
• A membrane keypad with alphanumeric keys
• Adjustable plate shake frequency and times
• Reads 96-well microplates with 0.355" (9.017 mm) well centers
• Internal, four-voltage range power input module that can be adjusted for 100, 120,
230, or 240 V~ @ 50-60 Hz (readers manufactured before December 2005)
• 24-volt external power supply compatible with 100-240 V~ ± 10% @ 50-60 Hz
(readers manufactured after December 2005)
• One serial COM port (25-pin male) and one parallel port (25-pin female)
• 4-Zone incubation chamber option (ELx808IU)

BioTek Instruments, Inc.

 Software Features | 3

Software Features

• Easy-to-use, menu-driven interface

• Endpoint, Kinetic, and Linear Well scanning calculations
• Curve fitting, with 4-parameter, cubic, quadratic, linear, 2-P, cubic-spline and
point-to-point methods
• Transformation and formula calculations for more complex mathematical
operations, including validation and cutoff formulas
• 55 assays are available onboard; up to 75 custom assays can be preprogrammed
and exported to the reader using BioTek’s Define Protocol software.
• Automatically stores results for the last 10 plates.

Package Contents

™ Package contents and part numbers are subject to change. Please

contact BioTek Customer Care with questions.

• ELx808 Absorbance Microplate Reader

• Power cord (PN varies according to country of use)
• Power supply for readers manufactured after December 2005 (PN 01281)
• Filter wheel with four standard filters: 405 nm, 450 nm, 490 nm, 630 nm and two
blank filters. The ELx808IU also includes a 340 nm filter.
• Operator’s Manual (PN 7341000)
• Serial cable (PN 75053)
• Dust cover (PN 7342066)

Optional Accessories

™ Accessory availability and part numbers are subject to change. Contact

BioTek Customer Care with questions, or visit www.biotek.com and
use the Accessories search tool.

• Additional filters (please inquire for availability):

¾ 340 to 630 nm, PN 3100XXX (XXX is wavelength in nanometers)
¾ 640 to 900 nm, PN 3404XXX (XXX is wavelength in nanometers)

ELx808 Operator’s Manual

4 | Chapter 1: Introduction

• Replacement lamp assembly (PN 3400508)

• Filter wheel plug (PN 3122037)
• Absorbance Test Plate (PN 9000547 or PN 7260522)
• ELx808 IQ/OQ/PQ Package (PN 7340543)
• Gen5 Software (visit biotek.com or contact your local dealer for details)
• Absorbance Liquid Test Solutions:
¾ BioTek QC Check Solution No. 1 (PN 7120779, 25 mL; PN 7120782, 125 mL)
¾ BioTek wetting agent (PN 7773002)
• Adapter to connect the reader to a USB-only printer (PN 75135)

Specifications

Microplates

• Standard 96-well, flat- or round-bottom plates

Electrical

• Power source and voltage range:

¾ Readers manufactured after December 2005: 24-volt external power
supply compatible with 100-240 V~ ± 10% @ 50-60 Hz
¾ Readers manufactured before December 2005: Internal, adjustable
power input module with four voltage ranges accommodated by the voltage
selection switch:
Range 1 100 V~ 90 to 110 V~, 50 to 60 Hz
Range 2 120 V~ 108 to 132 V~, 50 to 60 Hz
Range 3 230 V~ 207 to 253 V~, 50 to 60 Hz
Range 4 240 V~ 216 to 264 V~, 50 to 60 Hz
• Power consumption: 100 VA

Physical

• Dimensions: 16.0" D x 15.5" W x 8.75" H (40.6 cm x 39.37 cm x 22.2 cm)

• Weight: 35 lb. maximum (15.87 kg maximum)

BioTek Instruments, Inc.

 Specifications | 5

Environmental

• Operating temperature: 18° to 40°C (64° to 104°F)

• Humidity: 10% to 85% noncondensing

Hardware

• Light source: Tungsten halogen-filled bulb

• Display: 2-line x 24-character LCD

Reading Speeds

• Single wavelength: 8 seconds rapid mode; 12 seconds normal/regular mode

• Dual wavelength: 13 seconds rapid mode; 20 seconds normal/regular mode

• Single wavelength higher than 400 nm: 6-second minimum kinetic interval,
rapid mode

Standard Model: ELx808

™ The following specifications apply only to standard 96-well, flat- or

round-bottom microplates.

• Wavelength Range: 380 to 900 nm

• Filters: 10 nm half-bandwidth interference filters. User-accessible filter wheel.

Up to 6 filters may be installed on the instrument at one time. Filters supplied:
405 nm, 450 nm, 490 nm, 630 nm, and two blank filters.
• Absorbance Measurement Range: 0.000 to 4.000 OD

Optical specifications for single-wavelength endpoint measurements with a 12-

second read (normal/regular read mode):
Accuracy: ± 1.0% ± 0.010 OD from 0.000 to 2.500 OD @ 405 nm
Linearity: ± 1.0% from 0.000 to 2.500 OD at 405 nm
± 2.0% from 2.500 OD to 3.500 OD @ 405 nm
Repeatability: ± 0.5% ± 0.005 OD from 0.000 to 2.500 OD @ 405 nm
± 1.5% ± 0.005 OD from 2.500 to 3.500 OD @ 405 nm
± 2.5% ± 0.005 OD from 3.500 to 4.000 OD @ 405 nm

ELx808 Operator’s Manual

6 | Chapter 1: Introduction

Optical Specifications for single-wavelength kinetic measurements with read

intervals of less than 12 seconds:
Accuracy: ± 2.0% ± 0.010 OD from 0.000 to 2.500 OD @ 405 nm
Linearity: ± 2.0% from 0.000 to 2.500 OD @ 405 nm
Repeatability: ± 1.0% ± 0.010 OD from 0.000 to 2.500 OD @ 405 nm

™ The plate can be read at 6-second intervals (rapid mode) with

wavelengths higher than 400 nm.

Incubator/Ultraviolet Model: ELx808IU

™ The following specifications apply only to standard 96-well,

flat- or round-bottom microplates.

• Wavelength Range: 340 to 900 nm

• Filters: 10 nm half-bandwidth interference filters. User-accessible filter wheel.

Up to 6 filters may be installed on the instrument at one time. Filters supplied:
340, 405, 450, 490, 630 nm.
• Absorbance Measurement Range: 0.000 to 4.000 OD for 400 to 900 nm range,
0.000 to 3.000 OD for 340 to 400 nm range

Optical specifications for the 400 to 900 nm range (12-second read in normal/
regular read mode):
Accuracy, Linearity, Repeatability: Same as Standard Model.

Optical specifications for the 340 to 400 nm range (12-second read in normal/
regular mode):
Accuracy: ± 1.0% ± 0.010 OD from 0.000 to 2.000 OD @ 340 nm
Linearity: ± 1.0% from 0.000 to 2.000 OD @ 340 nm
Repeatability: ± 1.0% ± 0.005 OD from 0.000 to 2.000 OD @ 340 nm

• Incubation: The following specifications apply to a 96-well sealed plate with

200 μL of liquid in all wells:
¾ Temperature Control: Temperature controlled to 50°C

¾ Temperature Variation: ± 0.50°C @ 37°C with the plate sealed

BioTek Instruments, Inc.

 Product Support and Service | 7

Product Support and Service

If your instrument or software fails to function properly, if you have questions about how
to use or maintain your products, or if you need to send an instrument to BioTek for
service or repair, please contact our Technical Assistance Center (“TAC”).

Contacting the Technical Assistance Center

TAC is open from 8:30 AM to 5:30 PM (EST), Monday through Friday, excluding
standard U.S. holidays. You can send a fax or an e-mail any time.
Phone: 800-242-4685 (in the U.S.) or 802-655-4740 (outside the U.S.)
Fax: 802-654-0638
E-Mail: tac@biotek.com
Web: www.biotek.com

Please be prepared to provide the following information:

• Your name and company information
• A daytime phone or fax number, and/or an e-mail address
• The product name, model, and serial number
• The onboard software part number and version (available via the keypad
by selecting UTIL > TESTS > CHKSUM)
• Gen5 software version information (Help > About Gen5).

Returning Instruments for Service/Repair

If you need to return an instrument to BioTek for service or repair, please contact the
TAC for a Return Materials Authorization (RMA) number and the shipping address.
Repackage the instrument according to the instructions at the end of Chapter 2:
Installation.

ELx808 Operator’s Manual

8 | Chapter 1: Introduction

BioTek Instruments, Inc.

Chapter 2

Installation

This chapter includes instructions for unpacking and setting up the

ELx808, and instructions for connecting printers and/or serial
devices.

Product Registration ......................................................... 10 

1: Unpack and Inspect the Instrument ................................ 10 
2: Select the Operating Environment .................................. 11 
3: Install the Filter Wheel .................................................. 12 
4: Check/Adjust the Power Input Voltage Setting ................. 14 
5: Connect Power ............................................................ 14 
6: (Optional) Connect Printer ............................................. 15 
Printers ...................................................................... 16 
7: Turn on Reader and Run System Test ............................. 16 
8: Check/Adjust the Reader’s Filter Table ............................ 17 
9: Configure Reader Settings ............................................. 18 
SETUP Options ............................................................ 18 
OUTPUT Options .......................................................... 19 
REPORT Type .............................................................. 19 
READ Options ............................................................. 20 
Read Speed ................................................................ 21 
10: Install Software/Connect to Computer (Optional) ............ 21 
Attach the Cable ......................................................... 21 
Install Gen5 Software on the Host Computer ................... 21 
Establish Communication .............................................. 22 
Changing the Baud Rate on the ELx808 .......................... 22 
11: Verify Performance ..................................................... 23 
Before Repackaging the Instrument .................................... 24 
10 | Chapter 2: Installation

Product Registration
Please register your product(s) with BioTek to ensure that you receive important
information and updates about the product(s) you have purchased. Register online
through BioTek’s Customer Resource Center (CRC) at www.biotek.com or by contacting
BioTek Customer Care.

1: Unpack and Inspect the Instrument

Important! Save all packaging materials! If you need to

ship the reader to BioTek for repair or replacement, you must
use the original packaging materials. Using other forms of
commercially available packaging materials is not
recommended and can void the warranty. Improper
packaging that results in damage to the instrument may lead to
additional charges.

If the original packaging materials have been damaged or lost,

contact BioTek and ask for PN 7343000. See Product Support
& Service in Chapter 1 for contact information.

See Before Repackaging the Instrument at the end of this

chapter for complete shipping instructions.

™ The instrument’s packaging is subject to change. If the instructions in this

section do not apply to the packaging materials you are using, please contact
BioTek’s Technical Assistance Center for guidance.

The ELx808 and its accessories are securely packaged inside custom-designed shipping
materials. This packaging should protect the instrument from damage during shipping.
Inspect the shipping box, packaging, instrument, and accessories for signs of damage.
If the shipping box has been damaged: Inspect the instrument for visible dents and
scratches as you unpack it.
If the reader is damaged: Notify the carrier and your BioTek sales representative.
Keep the shipping cartons and the packing materials for the carrier’s inspection. BioTek
will arrange for repair or replacement of your reader immediately.
1. (Refer to Figure 1 on the next page.) Carefully open the top of the box, and
remove any accessories. These include a power cord and power supply, a filter
wheel in a padded envelope, and an operator’s manual.
2. Remove the end caps from the reader.

BioTek Instruments, Inc.

 2: Select the Operating Environment | 11

3. Lift the reader out of the box, and place it on a level surface. Remove the reader
from the plastic bag.
4. Remove the filter wheel from the shipping envelope
5. Place all packing material back into the shipping box for reuse if the instrument
needs to be shipped again.

Figure 1: Unpacking the ELx808

2: Select the Operating Environment

For best operation, install the reader on a level surface in an area where ambient
temperatures between 18°C (64°F) and 40°C (104°F) can be maintained.
The reader is sensitive to extreme environmental conditions. Conditions to avoid are:
• Excessive humidity: Condensation directly on the sensitive electronic
circuits can cause the instrument to fail internal self-checks. The humidity must
be in the range of 10% to 85%, noncondensing.
• Excessive ambient light: Bright sunlight or strong incandescent light can
reduce the linear performance range and affect the instrument’s readings.

ELx808 Operator’s Manual

12 | Chapter 2: Installation

• Dust: Optical density readings may be affected by extraneous particles (such

as dust) in the microplate wells. A clean work area is necessary to ensure
accurate readings.

3: Install the Filter Wheel

The filters that ship with the ELx808 are installed in the six-position filter wheel. For
example, the standard models have 405, 450, 490, 630 nm filters; the UV model’s filter set is
405, 450, 490, 630 and 340 nm.
The filter wheel, which is packaged in a shipping envelope, must be installed before the
reader is used. If you plan to install additional filters, or change the filter locations, use the
following instructions to gain access to the filter wheel (refer to

Figure 2 on page 13):

1. If the reader is on, turn it off and disconnect the power cord or power supply.
2. Remove the seven screws around the perimeter of the shroud with a
screwdriver.
™ Tip: Bring the reader to the edge of the work surface to access the screws
without having to turn the reader upside down.
3. Carefully lift up the shroud from the front. (Note that the shroud is hinged along
its back edge.) Hold the plate access door steady, or tape it closed as the shroud
is being lifted to prevent the door from moving.
4. Ensure that all locations on the filter wheel contain either a filter or a blank. Each
location on the filter wheel must be occupied for the reader to operate properly.
Take a moment to record the filter values in each location
(e.g., 405 nm in position 1).

Important! Keep track of all filter locations. The physical

location of the filters must match the filter locations mapped in
the reader’s software filter table. The filter wheel must have no
empty locations; all locations must be filled with either a filter
or a blank plug. Install all filters with the arrow denoting the
light direction pointing downward.

5. The filter wheel mount is located in the left-rear corner inside the reader. The
filter wheel is held in place by a magnet on the filter wheel motor hub. To install
the filter wheel:
• Line up the registration notch on the hub and the corresponding peg on the
filter wheel.
• Apply firm pressure to attach the filter wheel to the hub. The peg must be
engaged in the notch for proper installation.

BioTek Instruments, Inc.

 3: Install the Filter Wheel | 13

• Ensure that the filter wheel is positioned flat against the hub and that it
rotates freely.
• Lower the top shroud back into position and remove tape if used to steady
the plate access door.
• Reinstall the seven screws removed from the perimeter of the shroud.

Important! Be sure to replace the seven perimeter screws;

they increase the reader’s ability to withstand electrostatic
discharges and electromagnetic interference. In addition, the
screws MUST be installed to hold the shroud in place if you
plan to ship the instrument back to BioTek for service or repair.

6. To remove the filter wheel, grasp the center hub of the filter wheel and pull it
toward the lamp. The wheel should easily disconnect from the hub. Lift the
wheel from the instrument.
7. To properly store interference filters for the ELx808 during extended periods of
non-use, package the filters in a light-tight envelope or container, away from
high humidity and direct sunlight. This will ensure the longest life for the filters.
When handling the filters, keep the surfaces clean from fingerprints and debris
by simply wiping with a lens tissue or other lint-free cloth.

Figure 2: Installing the Filter Wheel

ELx808 Operator’s Manual

14 | Chapter 2: Installation

4: Check/Adjust the Power Input Voltage Setting

For ELx808 readers manufactured before December 2005 only: The reader is
powered by an internal, adjustable four-voltage range power input module, instead of by
an external power supply. For instructions for checking or adjusting the power input
voltage setting, and for reconfiguring or replacing the fuses, refer to Appendix C,
Adjusting the Power Input Voltage Setting.

5: Connect Power

ELx808 with the external power supply:

1. Connect the power cord to the external power supply.
2. Locate the power inlet on the right side of the reader.
3. Plug the rounded end of the power supply’s line cord into the power inlet.
4. Plug the other end of the power cord into an appropriate power receptacle.

ELx808 with the internal power input module:

1. Locate the power inlet on the right side of the reader.
2. Plug the rounded end of the power cord into the power inlet.
3. Plug the other end of the cord into an appropriate power receptacle.

Warning! Power Rating. The ELx808 or power supply

must be connected to a power receptacle that provides voltage
and current within the specified rating for the system. Use of
an incompatible power receptacle may produce electrical
shock and fire hazards.

Warning! Electrical Grounding. Never use a plug adapter

to connect primary power to the ELx808 power supply. Use of
an adapter disconnects the utility ground, creating a severe
shock hazard. Always connect the power supply cord directly
to an appropriate receptacle with a functional ground.

BioTek Instruments, Inc.

 6: (Optional) Connect Printer | 15

6: (Optional) Connect Printer

If the ELx808 will operate in standalone mode (that is, without BioTek’s Gen5 software
running on a host PC), connect a printer directly to the reader using the supplied cable.

Important! To avoid system instability, make sure the printer

and reader are turned OFF before connecting them.

1. If the reader and/or printer are on, turn them off. Place the printer in a location
adjacent to the reader.
2. Attach one end of the parallel cable to the printer’s parallel port.
3. Attach the other end of the cable to the reader’s parallel port, located on the
instrument’s rear panel.
4. Make sure the securing screws on both ends of the cable are tightened.
5. Turn on the printer.

Figure 3: Serial and parallel ports

ELx808 Operator’s Manual

16 | Chapter 2: Installation

Printers

The ELx808’s parallel port (LPT1) allows connection to compatible printers. The reader
supports printers using either HP's PCL3 language, such as the HP DeskJet series, or
Epson's LQ language. For a list of compatible printers call BioTek's Technical
Assistance Center or visit our website (refer to Chapter 1 for contact information).

7: Turn on Reader and Run System Test

If all of the required steps preceding this one have been performed successfully, locate the
reader’s power switch (on the right side) and turn on the reader. The ELx808 will
automatically perform a System Test. The System Test conducts a series of tests at each
wavelength defined in the filter table to confirm adequate light levels, low electronic noise,
adequate photodiode sensitivity, overall system cleanliness, and (if equipped) proper
function of the incubator. The testing verifies that the reader will give in-specification
performance for each set wavelength over the specified OD range.
If a printer is connected to the reader, the test results automatically print. This test can also
be run through Gen5 running on a host PC; consult the Gen5 Help system for instructions.
• If the test passes, the reader will beep once and the display will show the
software’s main menu, which will resemble one of the following:
Standard ELx808:

R E A D Y 0 1 : 3 0 P M 1 0 / 0 9 / 1 0
R E A D D E F I N E R E P O R T U T I L

ELx808IU:

R E A D Y 0 1 : 3 0 P M 3 7 . 0 º C
R E A D D E F I N E R E P O R T U T I L

• If the test fails, the reader will beep repeatedly and the display will show an
error code. If this happens, write down the error code and then press the STOP
key on the keypad to stop the beeping. Look up the error code in Chapter 6,
Error Codes to determine its cause.

If the problem is something you can fix (for example, if the error code is 0201
or 0301, indicating that the filter wheel is missing), turn off the reader, fix the
problem, and then turn the reader back on. If the cause is not something you
can fix, contact BioTek’s Technical Assistance Center. See Chapter 1,
Introduction for contact information.

BioTek Instruments, Inc.

 8: Check/Adjust the Reader’s Filter Table | 17

8: Check/Adjust the Reader’s Filter Table

After installing the filter wheel (or new filters), ensure that the ELx808’s filter table
accurately maps the physical location of the filters in the filter wheel.
1. Turn on the reader if it is not already on.
2. At the Main Menu screen, select UTIL.

R E A D Y 0 1 : 3 0 P M 1 0 / 0 9 / 1 0
R E A D D E F I N E R E P O R T U T I L

The SELECT UTILITY OPTION menu appears:

S E L E C T U T I L I T Y O P T I O N :
T E S T S S E T U P O U T P U T R E A D

3. From the Select Utility Option menu, select SETUP. The EDIT SETUP
INFORMATION screen appears:

E D I T S E T U P I N F O R M A T I O N :
D A T E T I M E F I L T E R * M O R E

4. Select FILTER. The wavelength currently defined for Filter #1 appears:

E N T E R
F I L T E R # 1 W A V E L E N G T H : 4 0 5

If you need to change the filter wavelength number, use the numeric keypad to
enter a number at the cursor location. To save the entry and move to the next
filter on the filter table, press the Enter key. When the last filter has been
entered, the software exits the filter routine, and displays the following screen:

E D I T S E T U P I N F O R M A T I O N :
D A T E T I M E F I L T E R * M O R E

5. Press the Main Menu key to return to the Main Menu.

ELx808 Operator’s Manual

18 | Chapter 2: Installation

9: Configure Reader Settings

The ELx808 may be configured a number of ways, depending on user preference.
Configuration options are accessed via the Select Utility Option menu, and include:
• SETUP: Set the date and time.

• OUTPUT: Select whether reports will be output to a Printer, the Computer, or

both, choose the Column or Matrix report format, and determine if Curves will
be printed.
• READ: Indicate whether prompts shall be displayed at run-time for Plate IDs,
Sample IDs, and Sample Counts.
• TESTS: Access functions to test reader optics and instrument calibration.
Check the version of basecode software installed. See Chapter 4,
Instrument Qualification for more information.

To select the SETUP, OUTPUT, and READ user-configurable options:

• At the Main Menu screen, select UTIL to access the Select Utility Option menu.

S E L E C T U T I L I T Y O P T I O N :
T E S T S S E T U P O U T P U T R E A D

SETUP Options

1. At the Select Utility Option screen, select SETUP.

E D I T S E T U P I N F O R M A T I O N :
D A T E T I M E F I L T E R * M O R E

2. At the Edit Setup Information screen, select DATE.

D A T E : 1 0 / 0 9 / 0 7 M D Y
M M D D Y Y D D M M Y Y

3. Enter the new date, using the numeric keys. The cursor is positioned under the
first editable field, and advances automatically. To change the date format, select
MMDDYY or DDMMYY. The display updates to reflect the new format.
4. Press Enter to return to the Edit Setup Information screen.

T I M E : 0 3 : 1 1 P M 1 2 H R
1 2 H O U R 2 4 H O U R A M / P M

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 9: Configure Reader Settings | 19

5. To edit the Time, select TIME. At the Time Entry Screen, use the numeric keys to
enter the correct time.
6. Select a 12- or 24-hour format by pressing the soft key beneath these options. The
display automatically updates with the new time AM/PM format.
7. Press the Previous Screen key to return to the Select Utility Option menu.

OUTPUT Options

1. At the Select Utility Option screen, select OUTPUT to set report output
preferences.

R E P O R T O U T P U T ? B O T H
P R I N T C O M P U T E R B O T H

2. Any previously defined selection appears on the top line of the display. Select
the desired option (PRINT, COMPUTER, BOTH) to change the output device.
3. Press Enter to advance to the Select Printer menu screen. If COMPUTER is
selected, all results will be transferred directly to the computer screen via the
RS232 serial port.

S E L E C T P R I N T E R E P S O N
E P S O N H P

REPORT Type

™ These selections only apply when the reader is run in standalone mode
(without Gen5). See Appendix A for examples of Reports.

1. At the Select Printer screen, press Enter to advance to the REPORT TYPE menu
screen.

R E P O R T T Y P E : M A T R I X
C O L U M N M A T R I X BO T H

2. The current selection appears on the top line of the display. Select the desired
option (COLUMN, MATRIX, BOTH) to change the Report Type.

ELx808 Operator’s Manual

20 | Chapter 2: Installation

3. Press Enter to advance to the SAMPLES ON COL RPT screen.

S A M P L E S O N C O L R P T ? N O
Y E S N O

4. Select YES to print samples on the column report, or NO to omit them.

5. Press Enter to advance to the PRINT CURVE-FIT screen.

P R I N T C U R V E - F I T ? N O
Y E S N O

6. The current selection is displayed on the top line of the screen. To change the
report option, select YES or NO. The display updates to reflect the selection.
(Select YES if there are quantitative assays defined on board and you wish to
print the curve.)
7. Press Enter to return to the Select Utility Option screen.

READ Options

1. At the Select Utility Option screen, press the soft key beneath READ to set up
Reader Prompt preferences. Select YES to prompt the operator to enter
identifications and sample counts before a microplate is read. The prompts are
shown below.
2. Press Enter after each selection to advance the display.

P R O M P T F O R P L A T E I D ? N O
Y E S N O

3. PROMPT FOR PLATE ID allows the user to enter an alphanumeric name of up

to 10 characters. Select YES to present this prompt at run-time.

P R O M P T F O R S A M P L E I D ? N O
Y E S N O

4. PROMPT FOR SAMPLE ID allows the user to identify samples with a

4-character alphanumeric name. The starting ID is entered and automatically
incremented by the software. Select YES to present this prompt at run-time.

P R O M P T S A M P L E C O U N T ? N O
Y E S N O

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 10: Install Software/Connect to Computer (Optional) | 21

5. PROMPT SAMPLE COUNT allows the user to enter the total number of samples
on the plate at runtime; only those sample results will be printed. Select YES to
present this prompt at run-time.

Read Speed

R E A D I N R A P I D M O D E ? N O
Y E S N O

RAPID MODE reads a 96-well plate at 6-second intervals with wavelengths higher
than 400 nm. Selecting NO means a plate with single-wavelength kinetic
measurements will be read in intervals less than 12 seconds (“Regular” or “Normal”
Mode). Specifications for these modes are outlined in Chapter 1.

10: Install Software/Connect to Computer

(Optional)
The ELx808 has serial (RS232) port located on back of the reader (see Figure 3 on page 15).
This port allows the reader to communicate with a computer using the BioTek Gen5
software. It also allows for upgrades to the ELx808 basecode (on-board) software.

Attach the Cable

• Turn off the computer and the reader.

• Connect the supplied serial cable to both machines.

Install Gen5 Software on the Host Computer

If applicable, install Gen5 on the host computer. There is a

certain sequence of events that must be followed to ensure
that the software is properly installed and configured. Please
follow the instructions provided in Gen5 Getting Started
Guide to install the software.

ELx808 Operator’s Manual

22 | Chapter 2: Installation

Establish Communication

1. Start Gen5 and log in if prompted. The default System Administrator password is
admin.
2. Go to the Gen5 main screen:
• Gen5 version 2.x users: From the Task Manager, select Setup > Go to System
Menu.
• Gen5 version 1.x users: From the Welcome screen, select System Menu.
3. Select System > Instrument Configuration and click Add.
4. Set the Reader Type to ELx808.
5. Set the Com Port to the computer’s COM port to which the reader is connected.
6. Click Test Comm. Gen5 attempts to communicate with the reader. If the
communication attempt is successful, return to the Gen5 main screen.
If the communication attempt is not successful, try the following:
• Is the reader connected to the power supply and turned on?
• Is the communication cable firmly attached to both the reader and the
computer?
• Did you select the correct Reader Type in Gen5?
• Try a different COM port.
• Make sure the reader display is at its Main Menu.

If you remain unable to get Gen5 and the reader to communicate with each other,
contact BioTek’s Technical Assistance Center.

Changing the Baud Rate on the ELx808

™ Gen5 requires the baud rate to be set to 9600.

If you need to change the baud rate from the default of 9600 to either 1200 or 2400:
1. Turn on the instrument if it is not already on.

R E A D Y 0 1 : 3 0 P M 1 0 / 0 9 / 0 7
R E A D D E F I N E R E P O R T U T I L

2. At the Main Menu screen, select UTIL.

S E L E C T U T I L I T Y O P T I O N :
T E S T S S E T U P O U T P U T R E A D

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 11: Verify Performance | 23

3. At the Select Utility Option screen, select SETUP.

E D I T S E T U P I N F O R M A T I O N :
D A T E T I M E F I L T E R * M O R E

4. At the Edit Setup Information screen, select *MORE to advance to the EDIT
SETUP / RS232 option screen.

E D I T S E T U P I N F O R M A T I O N :
R S 2 3 2 C A L P L A T E * M O R E

5. Select RS232 to access the Select Baud Rate menu. The top line of the
display shows the baud rate currently set.

S E L E C T B A U D R A T E : 9 6 0 0
1 2 0 0 2 4 0 0 9 6 0 0 V I E W

6. To change the Baud rate, press the soft key beneath the desired baud rate.
The display (top line) automatically updates to reflect the new choice.
7. To view the reader’s settings for parity, stop bits, and data bits, select
VIEW.

11: Verify Performance

Before using the ELx808 for the first time, verify that the reader is operating properly by
running a System Test and the Absorbance Plate Test. These tests and additional
verification procedures are described in Chapter 4, Instrument Qualification.

ELx808 Operator’s Manual

24 | Chapter 2: Installation

Before Repackaging the Instrument

Important! Use the instrument’s original shipping container

and packaging material. This shipping system was designed to
be used no more than five times. If the container is damaged
and/or has been used more than five times, contact BioTek for
a new set of shipping materials, and ask for PN 7343000.

The instrument’s packaging design is subject to change. If the

instructions in this section do not appear to apply to the
packaging materials you are using, please contact BioTek’s
Technical Assistance Center for guidance.

Warning! If the reader has been exposed to potentially

hazardous material, decontaminate it to minimize the risk to
all who come in contact with the reader during shipping,
handling, and servicing. Decontamination prior to shipping is
required by U.S. Department of Transportation regulations

1. Decontaminate the reader prior before shipping. Refer to Decontamination in

Chapter 5, Preventive Maintenance, for instructions.
2. Once the reader is clean, pack it in its original shipping box, using the original
packing materials (see Figure 1 on page 11).
3. Contact BioTek’s Technical Assistance Center for a Return Materials
Authorization (RMA) number and shipping instructions.

BioTek Instruments, Inc.

Chapter 3

Operation

This chapter includes instructions for operating the ELx808 Reader

and its software.

Getting Started with Gen5 ................................................. 27

Introduction .................................................................... 28
The Keypad ................................................................ 29
The Startup Screen...................................................... 30
The Main Menu Screen ................................................. 30
Quick Read ................................................................. 31
Defining Assays ............................................................... 32
Selecting an Assay ...................................................... 32
Assay Name ............................................................... 32
Defining the Method, Map, Formula, and Curve ................ 33
METHOD .................................................................... 34
MAP .......................................................................... 40
FORMULA ................................................................... 53
CURVE ....................................................................... 64
Panel Assays .............................................................. 67
Reading a Microplate ........................................................ 70
Select Assay ............................................................... 70
Run-Time Prompts ....................................................... 70
Enter Number of Samples ............................................. 71
Enter Plate ID ............................................................. 71
Enter Sample ID ......................................................... 71
Prompts for Well Location ............................................. 72
26 | Chapter 3: Operation

Beginning the Plate Read .............................................. 72

Printing Reports and Assay Lists......................................... 73
Result ........................................................................ 74
Editing Standard Outliers .............................................. 74
Printing Results ........................................................... 75
Map........................................................................... 75
Assay ........................................................................ 76
List ........................................................................... 76
Recommendations for Optimum Performance ....................... 76

Some readers have custom programmed assays installed.

Not all features of the software discussed in this chapter are
available on custom instruments. Please contact BioTek’s
Technical Assistance Center (TAC) if you have any questions
about the assays on your reader

All users should read Recommendations for Optimum

Performance on page 76.

BioTek Instruments, Inc.

 Getting Started with Gen5 | 27

Getting Started with Gen5

These instructions describe how to create and run an Experiment in Gen5. For more
information, or if the instructions below do not match what you see in Gen5, refer to the
Gen5 Getting Started Guide and help system.
Gen5 version 2.x users:
1. Start Gen5.
2. If the Task Manager appears, select Read Now > New and skip to step 4.
Otherwise, select File > New Task from the main view.
3. Select Read Now > New. Gen5 will open the procedure dialog. Go to step 4.
Gen5 version 1.x users:
1. Start Gen5. If the Welcome screen appears, select Read a Plate and skip to
step 4. Otherwise, select File > New Experiment from the main view.
2. Click Default Protocol and click OK. Gen5 will open the Experiment
workspace, which includes the Protocol menu tree and Plate screen.
3. Select Plate > Read or click the Read Plate icon. The Procedure dialog will
open. Go to step 4.
For any version:
4. Select a Plate Type.
5. Click Read to open the Read Step dialog.
6. Select a Read Type.
7. Select or enter the wavelength(s) at which the plate will be read.
8. Define other reading parameters as desired. Click Help for assistance.
9. When complete, click OK to return to the Procedure dialog.
10. Click OK to save and close the Procedure dialog.

• Gen5 version 1.x only: The Plate Reading dialog will open. Enter any
desired information, place the plate on the carrier, then click READ to
begin the plate read. If the Save As dialog opens, enter a File name,
choose a file location (Save in:) and click Save.
11. Click OK when the Load Plate dialog appears. The plate will be read.

• To view the raw data results, use the Data drop-down arrow in the
Plate screen to select one wavelength. The results will be displayed for
the selected wavelength. Repeat, for other wavelengths.
• To analyze, manipulate, or print results, Protocol parameters should be
defined. Refer to the Gen5 Help system for instructions.

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28 | Chapter 3: Operation

Introduction
The front panel on the ELx808 Reader features a 25-pad keypad and a 2-line x 24-character
LCD display as the user interface (see Figure 4 below). The reader’s bidirectional serial
port allows computer control of the instrument and provides the means for downloading
additional assay protocols to the instrument.

Figure 4: ELx808 Front Panel with LCD and Keypad

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 Introduction | 29

The Keypad

Use the four “soft keys,” located directly below the display, to select
options presented on the display. For example, from the Main Menu,
press the leftmost soft key to select READ, the rightmost to select UTIL.

Main Exit the current screen and return to the Main Menu. After defining a
Menu reader program, press the Main Menu key, then press YES to save the
program.

To scroll through the different options within a program, press the

Options Options key or the Shift + Options key combination. Press the
Enter key to select the current option.

Pressing Enter generally saves the current screen settings and

ENTER advances to the next screen in a series.

Previous To move to a previous menu, press the Previous Screen key.

Screen

Press Clear to clear/reset the current value on the reader’s LCD

CLEAR
display.

Press the ◄ (reverse) arrow to move the cursor to the left in the LCD
◄
display.

Press the ► (forward) arrow to move the cursor to the right in the LCD
► display.

READ To start running a reader program, press the Read key.

STOP To stop running (abort) a reader program, press the Stop key.

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30 | Chapter 3: Operation

The Startup Screen

To turn on the ELx808, press the on/off switch on the right side of the reader’s
base. The ELx808 will perform a system self-test, displaying the screens shown
below until initialization is complete. During this time, all keys are inactive.
If the self-test fails, the reader will “beep” and display an error code. Press Stop to
stop the beep. Refer to Chapter 6, Error Codes, to interpret these codes. Contact
BioTek Instrument’s Technical Assistance Center (see page 7), for further
assistance with troubleshooting.

P O W E R U P S E Q U E N C E V X . X X
I N I T I A L I Z I N G . . .

B I O T E K E L X 8 0 8
S E L F - T E S T . . . . .

The Main Menu Screen

Following successful power-up of the ELx808, the Main Menu screen is displayed.
This screen will vary slightly if the instrument has the Incubation/UV option:
Main Menu screen – ELx808

R E A D Y 0 1 : 3 0 P M 1 0 / 0 9 / 0 7
R E A D D E F I N E R E P O R T U T I L

Main Menu screen – ELx808IU (model with incubation/UV capability)

R E A D Y 0 1 : 3 0 P M 3 7 . 0 º C
R E A D D E F I N E R E P O R T U T I L

™ Note: The temperature indicated on the display of incubated models is

the actual averaged temperature of the incubator’s four zones. The
applied setpoint of the last assay is used. To adjust the temperature, a
new setpoint value must be assigned to the assay before running.

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 Introduction | 31

The keyboard’s four “soft keys,” located below the on-screen menu options
(READ, DEFINE, REPORT and UTIL), are activated, and may be selected. Press
the soft key that corresponds to a displayed menu option to select that option:
• READ option: Initiate a plate read (or, press the key labeled READ on the
keyboard for plate reading prompts). You will be prompted to select from a
list of available assays.
• DEFINE option: Create a reading and data reduction protocol. You will be
prompted to select/edit an assay and then define its various parameters.
• REPORT option: Print results reports and protocol descriptions. For results
reports, you will be prompted to select a previously run assay with valid
data.
• UTIL option: Access various onboard utilities, used for configuring and
testing the reader.

Quick Read

On some readers, Assay 01 has been designed to allow for quick and simple
assay programming. It appears as _Quick Read on the display. Most of the
options available in Assays 2 through 55, and described in this section, are
unavailable for programming within Quick Read. You can access the Quick Read
assay when READ is selected from the main menu.
The Quick Read assay DEFINE settings are shown below, and cannot be edited,
except where noted:

METHOD

• Single Wavelength 405 nm (editable)

MAP

• 96-well plate geometry

• Blank on Air
• Automap
• Map starting location A1
• Samples only (no blanks, standard, or controls)
• Sample count prompted at runtime (can be turned off in UTIL > READ
options)

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32 | Chapter 3: Operation

Defining Assays

™ Note: Appendix B, Instructions for Programming a New Assay,

provides two examples of assay kit instructions and step-by-step
directions for programming each assay. The appendix includes two
sample assays: one with a ratio transformation calculation and a
POS/NEG cutoff determination, and another with a standard curve.

The Main Menu option, DEFINE, allows you to customize previously defined assays
stored in the reader’s memory.
• From the Main Menu, press the soft key beneath the DEFINE menu option to
advance to the SELECT ASSAY NUMBER screen.

Selecting an Assay

At the SELECT ASSAY NUMBER screen:

1. Use the NUMERIC keys to enter the number of predefined Assay Definition
files stored in the reader’s memory, or use the OPTION key to advance one
assay at a time. The cursor is positioned at the first editable field, and
advances automatically. The numeric range depends on the number of
assays (1-55 or more if custom programmed) programmed in the reader’s
memory.
2. The assay’s name and number are displayed on the screen.

S E L E C T A S S A Y N U M B E R : 6 5
N A M E : H B S - A G 1

3. Press Enter to select the assay and advance to the Name screen. You may
change the default assay name to a more descriptive one (see Assay Name
on the next page).
4. Press Enter to advance to the Edit Assay Name screen.

Assay Name

At the Assay Name screen, edit the name assigned to the Assay. The assay name
can be up to 16 characters.

E D I T > H B S - A G 1
- / : S P A C E

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 Defining Assays | 33

• Press the ALPHA and NUMERIC keys to update the assay name. The cursor
is positioned at the first editable field.
• Press the OPTION key to sequentially advance the character positioned
above the cursor. The characters will cycle through the alphabet (A-Z), with
a space character following Z.
• Press Clear to clear the assay name from the display.
• Use the Left and Right Arrows to move the cursor to the previous or next
editable field. The cursor will wrap around the edit field.
• Press Soft Keys 1, 2, 3, and 4 when using alphanumeric characters on the
display above the soft key in the assay name.
In addition:
• Press the Main Menu key to return to the Main Menu screen.
• Press Previous Screen to save the contents of the display and return to
the previous screen.
• Press Enter to save the contents of the display and advance to the next
screen.

Defining the Method, Map, Formula, and Curve

The DEFINE Option screen allows you to edit the Method, Map, Formula or Curve
Fit.

D E F I N E
M E T H O D M A P F O R M U L A C U R V E

Press the soft key beneath the displayed option to access the following functions:
• SOFT KEY 1: METHOD is selected, and the user is prompted to select the
read method parameters.
• SOFT KEY 2: MAP is selected and the user is prompted for plate mapping
information.
• SOFT KEY 3: FORMULA is selected and the user is prompted to enter a
formula.
• SOFT KEY 4: CURVE FIT is selected and the user is prompted for curve-fit
options.
• In addition, the MAIN MENU, PREVIOUS SCREEN and ENTER keys are
active, allowing you to move back and advance through the menu
structure.

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34 | Chapter 3: Operation

METHOD

The definition of a method includes selecting:

• Endpoint, Kinetic or Scanning Read Modes
• Delay First Read
• Incubation Parameters
• Filter Wavelengths Applied
• Shake Parameters
• Kinetic Analysis

™ Note: Some screens shown below and on the following pages may not
appear on some reader models.

Read Type

This option allows you to enter which read type: Endpoint, Kinetic, or Scan.
The following keys are active during this screen:

R E A D T Y P E : K I N E T I C
E N D P O I N T K I N E T I C S C A N

• Press SOFT KEY 1 to select Endpoint read mode. Press SOFT KEY 2 to
select Kinetic read mode.
• Press Enter to save the displayed value and advance to the next screen.

Delay in First Read

Selecting the Delay in First Read option allows you to enter a time delay before
the first read.

D E L A Y F I R S T R E A D
T I M E : X X : X X

• Enter the time in minutes and seconds, using the numeric keys.
• Press Enter to advance to the INCUBATION TEMPERATURE screen.

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 Defining Assays | 35

Incubation Temperature

The incubation temperature screen allows you to set the assay incubation
temperature.

I N C U B A T I O N T E M P : 3 7 C
A M B I E N T T E M P E R A T U R E

• Press SOFT KEY 1 or 2 to select ambient incubation.

• Press SOFT KEY 3 or 4 to select an adjustable temperature.
• Use the LEFT and RIGHT ARROW keys to move the cursor between
the two digits on the input temperature.
• Use NUMERIC keys to enter the incubation temperature, up to 50°C.
• Press Enter to save and advance to the next screen.

Single or Dual Wavelength

The WAVELENGTH selection screen allows you to select SINGLE or DUAL

wavelength for the assay.

W A V E L E N G T H : D U A L
S I N G L E D U A L

• Press SOFT KEY 1 to select SINGLE wavelength. The reader will

measure the optical density of each well with a single filter.
• Press SOFT KEY 2 to select DUAL wavelength. Each well will be read
twice, each time with a different filter. Note: The microplate is not
removed from the reading chamber between the two measurements.
The final reported optical density is the difference between the two
readings. Dual wavelength readings can significantly reduce optical
interference caused by scratched or fingerprinted microplates since the
scratches or fingerprints reduce the amount of light on both
wavelengths.
• Press Enter to save the selection and advance to the next screen.

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36 | Chapter 3: Operation

MEAS Selection

The MEAS selection screen allows you to select the filter(s) for the assay. The
currently selected filter appears on the top line of the display, and the available
options appear on the bottom.

M E A S : 4 5 0 R E F : 6 3 0
4 0 5 4 5 0 4 9 0 6 3 0

• Press SOFT KEYS 1, 2, 3, and 4 to select the filter option displayed

above the soft key. The display updates to reflect the selection.
• Press the right arrow key to select the Reference Filter.
• Press ENTER to move to the next screen.

Number of Kinetic Reads/Kinetic Duration Selection

This menu allows you to either select the total number of kinetic reads or the
length of time the assay will run (kinetic duration). Any previously defined
value is shown on the top line of the display and the options on the second.

K I N E T I C : T O T A L R E A D S
T O T A L R E A D S D U R A T I O N

• Press SOFT KEY 1 to select the total reads option.

• Press SOFT KEY 3 to select the duration option.
• Press Enter to save the selection and advance to the next screen.

Kinetic Interval

Use this screen to enter the interval of time between each kinetic read.

K I N E T I C
I N T E R V A L : 0 1 : 2 3 : 5 6

• Use the NUMERIC keys to enter the time duration. Valid ranges are: 0-1
hours, 0-59 minutes and 0-59 seconds. The number of Reads =
Duration/Interval must be less than or equal to 40 and more than or
equal to 2.

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 Defining Assays | 37

• Use the LEFT and RIGHT ARROW keys to move to the next or previous
numeric entry fields.
• Press Enter to save the value and advance to the next screen.

Kinetic Number of Reads

T O T A L N U M B E R
O F K I N E T I C R E A D S : 0 6

• Use the NUMERIC keys to enter the number of reads required. The
range is 2 to 40 reads.
• Press Enter to save the entry and advance to the next screen.

Kinetic Duration

Use this screen to enter the duration of the kinetic reaction.

K I N E T I C
D U R A T I O N : 1 1 : 2 3 : 4 5

• Use the NUMERIC keys to enter the time duration in hours, minutes,
and seconds. The maximum duration time is 80 hours.
• Use the LEFT and RIGHT ARROW keys to move between entry fields.
• Press Enter to save the entry and advance to the next screen.

Shake Mode Selection

S H A K E : B E F O R E E V E R Y R E A D
F I R S T E V E R Y N O N E

• Press SOFT KEY 1 to select shaking for the first read only or endpoint
read.
• Press SOFT KEY 2 to select shaking for every read.
• Press SOFT KEY 3 to select no shaking.
• Press ENTER to save the selection and advance to the next screen.
• Press SCREEN key to save the selection and return to a previous screen.

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38 | Chapter 3: Operation

Shake Time

S H A K E T I M E : 0 0 : 1 2 : 3 4
C O N T I N U O U S

• Use the NUMERIC keys to enter the shake interval. Valid ranges are:
0-1 hours, 0-59 minutes and 0-59 seconds.
• Use the LEFT and RIGHT ARROW keys to move the cursor between
hours, minutes, and seconds.
• Press Enter to save the entry and advance to the next screen.

™ Note: "Continuous" appears on the display when a kinetic assay with a

previously specified shake has been selected.

Shake Speed

The shake movement is a repeated 0.021-inch movement from the shake

position and back.

S H A K E S P E E D : M E D I U M
L O W M E D I U M H I G H V A R I

• Press SOFT KEY 1 to select low-speed shaking.

• Press SOFT KEY 2 to select medium-speed shaking.
• Press SOFT KEY 3 to select high-speed shaking.
• Press SOFT KEY 4 to select variable-speed shaking (1 second of each
speed repeated).
• Press Enter to save the entry and advance to the next screen.

Kinetic Data Analysis Selection

K I N E T I C A N A L Y S I S : R - S Q R
R A T E R - S Q R O N S E T

• Press SOFT KEY 1 to select the kinetic rate calculation. This method
will apply a linear fit to calculate the maximum slope based on the
number of kinetic points specified.

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 Defining Assays | 39

• Press SOFT KEY 2 to select the R-squared rate calculation. This method
will calculate the R-squared value at the maximum slope, based on the
linear curve fit and the number of kinetic points specified.
• Press SOFT KEY 3 to select the time calculation, which will calculate
the time for each well to reach the onset optical density.
• Press ENTER to save the selection and advance.

Number of Kinetic Points Selection

Use this screen to select the number of sequential kinetic points to calculate the
steepest Rate, or the R squared at the steepest Rate.

K I N E T I C P O I N T S : 3
A L L P O I N T S

• Use the NUMERIC keys to input the number of points. The range is 2 to
MAX where max is the total number of reads.
• Press SOFT KEY 1 or 2 to select ALL POINTS.
• Press Enter to save the entry and advance to the next screen.

Onset OD Selection

Use this screen to enter the onset OD time.

E N T E R
O N S E T O D : 1 . 2 3 4

• Use NUMERIC keys to enter the onset OD. 3.000 OD is the maximum
value.
• Use the LEFT and RIGHT ARROW keys to move the cursor within the
entered OD field.
• Press the Enter key to save the entry and advance.

Linear Scanning

If Scanning is chosen as the Read Type, use the following screen to enter the
total number of points to be read in a line across the center of each well.

E N T E R N U M B E R O F
S C A N P O I N T S : 1 5

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40 | Chapter 3: Operation

The maximum number of points selectable is 31 (odd numbers only). The 31

scan positions are fixed in the software. You must determine the optimal
number of scans per well. If, for example, 7 scans across the well is chosen, the
reader will read the centermost seven points in the well. The more scan points
chosen, the closer to the well sides reads will be taken.

™ Note: If too many scans are chosen, the reader may be reading
the sides of the well.

The reader will read the chosen number of points across the well and report
the calculated area under the curve.

MAP

The MAP Definition screen allows you to edit or specify the following options in
the assay:
• Automatic or Manual Map Generation
• Mapping Direction
• Replication Direction
• Blank Map Selection
• Blanking Constant
• Number of Blanks
• Location of Blanks
• Number of Standards
• Number of Standard Replicates
• Averaging of Standards
• Concentration and Location of Standards
• Number of Controls
• Control Type Definition
• Number of Control Replicates
• Control Location
• Number of Samples
• Number of Sample Replicates
• Averaging of Samples
• Sample Location

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™ Note: Some of the following screens may not appear, depending on the
reader model. The valid range of the number of standards is 0-12. The
valid range of valid replicate counts for standards is 1-8. The valid range of
the number of controls is 0-8. The range of valid replicate counts for
controls and samples is 1-12.

D E F I N E :
M E T H O D M A P F O R M U L A C U R V E

• At the DEFINE Options screen, press SOFT KEY 2 to begin the plate MAP
process.

Map Generation

“Map Generation” represents the method by which blanks, controls, standards,

and/or samples are assigned to specific locations on the plate. The currently
selected value appears on the top line of the display, and the available options
appear on the bottom.

M A P G E N E R A T I O N : M A N U A L
A U T O M A N U A L

Automatic Plate Map Generation: Select AUTO to instruct the software to

automatically generate a plate map after the blanks, controls, standards,
and/or samples have been defined.
Manual Plate Map Generation: Select MANUAL to indicate that the well
assignments will be performed manually (by the user) at Define and/or Read
time.
• Press SOFT KEY 1 for automatic sample plate map generation. The
display will update to reflect the selection.
• Press SOFT KEY 2 for MANUAL plate map generation. The display
updates to reflect the selection.
• Press ENTER to save the selection and move to the next screen.

™ Note: Press SHIFT-CLEAR to clear any previously defined

manual map.

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42 | Chapter 3: Operation

Mapping Direction

Use this option to select how the wells are mapped on the plate. Any
previously defined Mapping Direction appears on the top line of the display;
the available options appear on the second line.

M A P P I N G D I R E C T I O N : D O W N
D O W N A C R O S S

• Press SOFT KEY 1 to map DOWN the column.

• Press SOFT KEY 2 to map ACROSS the row.
• Press ENTER to save the selection and move to the next screen.

Replication Direction

This option allows you to specify how replicates are mapped on the plate. The
currently selected Replication Direction appears on the top line of the display,
and the available options appear on the bottom.

R E P D I R E C T I O N : A C R O S S
D O W N A C R O S S

• Press SOFT KEY 1 to map the replicates DOWN the column, following
the direction of the map listing.
• Press SOFT KEY 2 to map the replicates ACROSS (in a paired format).
As an example, two replicates can be placed in A1 and A2 wells. The
third replicate would follow in B1. The next standard control, or
sample, would follow in B2.
• Press ENTER to save the selection and advance.

Examples of mapping and replication directions are shown on the

next page.

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 Defining Assays | 43

EXAMPLES OF MAPPING & REPLICATION DIRECTIONS

Map Direction DOWN, Rep Direction DOWN:

1 2 3 4 5 6 7 8 9 10 11 12
A STD1 STD5 SMP
B STD1 STD5 SMP
C STD2 PC SMP
D STD2 PC
E STD3 NC
F STD3 NC
G STD4 SMP
H STD4 SMP

Map Direction ACROSS, Rep Direction ACROSS:

1 2 3 4 5 6 7 8 9 10 11 12
A STD1 STD1 STD2 STD2 STD3 STD3 STD4 STD4 STD5 STD5 PC PC
B NC NC SMP SMP SMP SMP SMP SMP
C
D
E
F
G
H

Map Direction DOWN, Rep Direction ACROSS:

1 2 3 4 5 6 7 8 9 10 11 12
A STD1 STD1
B STD2 STD2
C STD3 STD3
D STD4 STD4
E STD5 STD5
F PC PC
G NC NC
H SMP SMP

Map Direction ACROSS, Rep Direction DOWN:

1 2 3 4 5 6 7 8 9 10 11 12
A STD1 STD2 STD3 STD4 STD5 PC NC SMP SMP
B STD1 STD2 STD3 STD4 STD5 PC NC SMP SMP
C
D
E
F
G
H

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Start Mapping at Well Location

The Start Mapping at Well Location screen is only shown if automatic mapping
is selected. This option allows you to enter the location of the well that will be
the starting point for automatic mapping.

S T A R T M A P P I N G
A T W E L L L O C A T I O N : A 0 1

• Use the LEFT and RIGHT ARROW keys to move the cursor to the
previous or next editable field. The cursor will wrap around the edit
field.
• Use the NUMERIC and ALPHA keys to enter a letter or number at the
cursor location. For all prompts of a well location, only the ALPHA keys
are active for the first character and NUMERIC for the second and third
characters.
• Press ENTER to save the well location and advance to the next screen.

Selecting a Blank Map

This option allows you to select which blanking method to apply to the assay.
The currently selected Blank Map value appears on the top line of the display,
and the available options appear on the bottom.
The blanking options, AIR, FULL and CONSTANT, ROW and COLUMN, and
P-ACROSS and P-DOWN are displayed on three screens.

B L A N K M A P : F U L L
A I R F U L L C O N S T * M O R E

B L A N K M A P : F U L L
R O W C O L U M N * M O R E

B L A N K M A P : F U L L
P - A C R O S S P - D O W N * M O R E

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• Press SOFT KEYS 1, 2, or 3 to select the BLANK MAP type above the
soft key. The display updates to reflect the selection.
• Press SOFT KEY 4 to access MORE options: ROW or COLUMN, and
P-ACROSS or P-DOWN.

• Press ENTER to save the well location and advance to the next screen.

Blank Map Definitions

• AIR performs an initial reading on “air” just prior to the plate read, and
uses that value as the blank value. This value is subtracted from each
well on the plate.
• FULL enables a single blank well or an average of blank wells to be
subtracted from the whole plate.
• CONST (Constant) allows entry of a user-specified absorbance value.
This value will be subtracted from each well on the plate. Use a blank
value from the first plate, or a blanking plate to save space on
subsequent assay plates.
• ROW enables a single blank well or an average of blank wells to be
selected for each row. The blank (or average) will be subtracted from
each well in the row. Use manual mapping to position blanks, controls,
standards, and samples.
• COLUMN enables a single blank well or an average of blank wells to be
selected for each column. The blank (or average) will be subtracted
from each well in the column. Use manual mapping to position blanks,
controls, standards, and samples.
• P-DOWN enables a blank in every even numbered column to be
subtracted from the well to the left of it in every odd column. Manual
mapping is recommended to set up the appropriate map by placing the
standards, controls, and samples in only the odd columns.
• P-ACROSS enables a blank in the B, D, F and H rows to be subtracted
from the well above in the A, C, E and G rows. Manual mapping is
recommended to set up the appropriate map by placing the standards,
controls, and samples in only the A, C, E, and G rows.

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46 | Chapter 3: Operation

Constant Blank Value Entry

This entry screen only appears when a Constant Blank map is selected. Enter
the value to be subtracted from each well on the plate.

E N T E R
B L A N K I N G C O N S T A N T : 0 . 0 0 0

• Use the NUMERIC keys to enter the value. The range is 0.000 to 3.000.
The cursor is positioned at the first editable field and advances
automatically.
• Press the CLEAR key to clear the value on the display.
• Press the ENTER key to save the value and advance.

Number of Blanks

The Number of Blanks field allows you to enter the number of blank wells in
the assay. This entry screen is only displayed when Full, Column, or Row
blank map is selected.

E N T E R N U M B E R O F
B L A N K S : 0 2

• Use the NUMERIC keys to enter the number of blanks. The range is 0 to
48.
• Press the CLEAR key to clear the Number of Blanks value from the
display.
• Press the ENTER key to save the value and move to the next screen.

Selecting a Blank Location

The Blank Location screen allows you to define where the blank well is located
on the microplate. This screen only appears if manual map generation has been
selected. Any previously defined value is displayed.

E N T E R T H E L O C A T I O N O F
B L A N K # 1 : A 1 2

• Use the NUMERIC and ALPHA keys to enter a Blank Location, based
upon the plate geometry.

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• Use the ARROW keys to move the cursor to the next or previous
editable field. The cursor is positioned beneath the first editable field.
• Press the ENTER key to save the value, and move to the next screen.

Number of Standards

Use this option to enter the number of standards that will be used in the assay.
Any previously defined value will be displayed on the screen. If the number of
standards is altered, the number of replicates for the standard automatically
defaults to 1.

E N T E R N U M B E R O F
S T A N D A R D S : 0 2

• Use the NUMERIC keys to enter the Number of Standards. The valid
range depends on the selected curve fit method. The maximum number
of standards is 12. The minimum is 4 for 4-P fit, cubic, cubic spline, and
2-P; 3 for quadratic; and 2 for linear and point-to-point.
• Press CLEAR to clear the value on the display.
• Press ENTER to save the value, and move to the next screen.

Number of Standard Replicates

E N T E R N U M B E R O F
S T A N D A R D R E P L I C A T E S : 0 2

• Use the NUMERIC keys to enter the Number of Replicates per

Standard. The range is 1 to 8 replicates. The software will verify that the
number of replicates, multiplied by the number of standards, does not
exceed the number of wells on the plate.
• Press CLEAR to clear the value on the display.
• Press ENTER to save the value and move to the next screen.

Average Standards

The Average Standards option allows you to select whether or not to average
the Replicates of a Standard. This average is used to calculate the standard
curve instead of using the individual replicate of each standard. Note: If the
replicate selection is 1, this option is not available.

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48 | Chapter 3: Operation

A V E R A G E S T A N D A R D S ? Y E S
Y E S N O

• Press SOFT KEY 1 to select YES (average the replicates). The top line
of the display updates to reflect the selection.
• Press SOFT KEY 2 to select NO (do not average the replicates). The
top line of the display updates to reflect the selection.
• Press the ENTER key to save the selection, and advance to the next
screen.

Standard Concentration

The Standard Concentration field allows you to enter the predicted or expected
concentration value for each standard group. Any previously defined value is
displayed.
If Manual Map Generation is selected, the replicate locations must also be
defined.

C O N C N O F L O C A T I O N
S T D 1 : 1 . 5 0 R E P # 1 : A 0 1

• Use the NUMERIC and ALPHA keys and the DECIMAL POINT key to
enter Standard Concentration values. The range is 0.00001 to 999999.
The entry cannot exceed six characters including the decimal point.
Valid well locations for the defined geometry are listed below.
• Use the RIGHT ARROW and LEFT ARROW keys to move to the next
or previous editable field.
• Press the CLEAR key to clear the Standard Concentration value from
the display.
• Press the ENTER key to save the value on the display and move to the
next screen.

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Reuse of Standard Curves

The ELx808 has the ability to reuse a standard curve that has already been
established.

Limitations of the Reuse of Standard Curves

• The reuse of standard curves can only be done in assay positions 31
through 55. Each of these positions can only store one standard curve.
• Standard curves cannot be reused on panels (see page 67 for Panel
Assays).

• Standard curves will be stored in memory with the Assay Name, Standard
Concentrations, Replicate Counts, and Optical Densities for each standard
replicate.
• Stored standard curves can only be reused for the assay that the curve was
originally generated on (i.e., the curve for assay #53 cannot be applied to
samples on a plate to be run in assay #51).
• To reuse a standard curve, an assay must first be programmed (in positions
31 through 55) and run. During the defining process, you will be prompted
to enter the number of standards, the number of standard replicates, and
the standard concentrations. The following screen has been added after
these prompts:

R E U S E S T A N D A R D C U R V E ? Y E S
Y E S N O

• After the assay has been run, the results have been calculated, and the
reports have been generated, the reader will prompt if this standard curve
should be stored in memory. The following display will appear:

S A V E S T A N D A R D C U R V E ? Y E S
Y E S N O

• Select YES to store the curve for use at a later time. The next time a plate is
to be read using this assay, the instrument will prompt if there are
standards on the plate. Select NO to discard the curve.

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50 | Chapter 3: Operation

S T A N D A R D S O N P L A T E ? N O
Y E S N O

• If YES is chosen, a new standard curve will be generated. The plate map is
not changed. (If “Prompt for Sample ID” is enabled in the UTIL section,
you will be prompted to enter the number of samples. See page 18 for more
information on the UTIL options.)
• If NO is chosen, the stored standard curve will be used. If Auto mapping
had been used to originally map the standards, blanks, controls, and
samples defined for this assay, the map will be automatically regenerated
without the standards, beginning in well xxx (where xxx was chosen as the
Starting well in the map, usually well A01). If Manual mapping was used to
map the plate, the map is NOT regenerated - the reader will NOT produce
results for the well positions that originally were standards. Auto mapping
is recommended, if the standards curves will be routinely reused.

Number of Controls

E N T E R N U M B E R O F
C O N T R O L S : 0 2

• Use the NUMERIC keys to enter the Number of Controls. The range
depends on the number of locations on the plate that are undefined.
The maximum number of controls is 8.
• Press CLEAR to clear the value on the display.
• Press ENTER to save the value and advance to the next screen.

Control Type

C O N T R O L # 1 : P C
P C N C H P C * M O R E

C O N T R O L # 1 : P C
L P C C T L 1 C T L 2 * M O R E

C O N T R O L # 1 : P C
C T L 3 C T L 4 * M O R E

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• Press the soft keys under the displayed Control Type to select the
option (Positive Control, Negative Control, High Positive Control, Low
Positive Control, CTL1, CTL2, CTL3, CTL4).

• Press CLEAR to clear the Control Type from the display.

• Press ENTER to save the displayed Control Type and advance to the
next screen.

Number of Control Replicates

E N T E R N U M B E R O F
R E P L I C A T E S O F P C : 0 2

• Use the NUMERIC keys to enter a value for Number of Control

Replicates. The range is 1 to 12 replicates. The software performs a
check to ensure the number of replicates does not exceed the number of
undefined wells remaining on the plate.
• Press CLEAR to clear the displayed Number of Replicates value.
• Press ENTER to save the displayed value and advance to the next
screen.

Location of Controls

The displayed location field can only be edited if manual map generation was
selected.

C O N T R O L # 1 L O C A T I O N
T Y P E : P C R E P # 1 : A 0 1

• Press the CLEAR key to clear the value on the display.

• Use the NUMERIC and ALPHA keys to enter values for well locations
on the plate.

Valid Well Locations

For all prompts of a well location, only the ALPHA keys are active for the first
character and NUMERIC for the second and third characters.
• Press the MAIN MENU and PREVIOUS SCREEN keys to move back
and advance through the menu structure.

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52 | Chapter 3: Operation

Number of Samples

E N T E R N U M B E R O F
S A M P L E S : 2 4

• Use the NUMERIC keys to enter the number of samples in the assay.
The range is 0 to the number of undefined well locations remaining on
the plate. If there are no controls, blanks, or standards defined on a 96-
well plate, the maximum number of samples is 96, and the minimum
number of samples is 1.
• Press the Clear key to clear values on the display.
• Press Enter to save the displayed value and advance to the next screen.

™ Note: If the number of samples is altered, the number of replicates for

the sample reverts to a value of 1.

Number of Sample Replicates

E N T E R N U M B E R O F
S A M P L E R E P L I C A T E S : 0 2

• Use the NUMERIC keys to enter the Number of Sample Replicates. The
range is 1 to 12 replicates. The software ensures that the number of
replicates multiplied by the number of samples, multiplied again by the
number of dilutions, does not exceed the number of undefined wells
remaining on the plate.
• Press the Clear key to clear the value on the display.
• Press the Enter key to save the displayed value and advance to the next
screen.

Sample Location

If Manual Map Generation is selected, this screen allows you to select the
well location of the sample on the plate. Any previously defined Sample
Location appears on the display.

S A M P L E # 1 L O C A T I O N
R E P # 1 : A 0 1

• Use the NUMERIC, ALPHA, and DECIMAL POINT keys to enter the
sample identifier and its location on the plate.

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• Press the CLEAR key to clear the displayed value.

• Press ENTER to save the value and advance to the next screen.

FORMULA

The ELx808 supports three types of formulas (Cutoff, Transformation, and Validation),
as well as the ability to define variables for use within formulas. Up to three types of
Validation formulas may be defined (Blank, Control, and Assay Validation).
Formulas are processed in the following order, with the number of permitted formulas
of each type:
• Blank Validation 0-1
• Control Validation 0-4
• Assay Validation 0-4
• Transformations 0-1
• Cutoff Formulas 0-1
• Curve Fit Analysis (if a curve fit method is defined)
Within any given formula type, the order of processing is the order in which the
formulas are entered.

Validation Formula Examples

• Blank Validation: An assay protocol states that every blank well on a

plate should have an OD of less than 0.050. The formula is entered on the
reader as a Blank Validation Formula: BLK < 0.050.
• Negative Control Validation: An assay protocol states that every
Negative Control well must have an OD of less than or equal to 0.100. The
formula is entered as a Control Validation Formula: NC < = 0.100.
• Positive Control Validation: An assay protocol states that every Positive
Control well must have an OD higher than 1.000, but less than 2.500. Two
Control Validation Formulas can be entered: PC > 1.000 AND PC < 2.500.
Or, one formula can be used if the formula is 24 characters or less: PC >
1.000 AND PC < 2.500.
• Assay Validation: An assay protocol states that in order for an assay to
be valid, the mean of the Negative Control well ODs must be less than
0.100. The Assay Validation formula that should be entered: NC;x < 0.100
(the map identifier NC;x indicates the mean of the NCs).
From the assay DEFINE Menu, press the arrow corresponding to FORMULA.

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Formula Type

The ELx808 supports three types of formulas, as well as the ability to program
variables for use within Transformation formulas.
• CUTOFF formulas are used to classify results. During data reduction,
results are evaluated against the cutoff formulas, and each well is
assigned a user-specified label or “call” (POS, NEG, or EQUIV).
• TRANSformation formulas are applied to the raw data in preparation
for further data reduction and/or curve-fit calculation.
• VALidation formulas can be used to determine whether or not blanks
and/or controls are valid. In addition, Assay Validation formulas can
be used to determine whether or not the entire assay should be
considered valid.
• The TRANS-VAR option allows you to define a variable to be used in
transformation formulas.

™ Note: GENERAL formulas are not used in the ELx808 open

assays.

S E L E C T F O R M U L A T Y P E :
C U T O F F T R A N S V A L * M O R E

S E L E C T F O R M U L A T Y P E :
G E N E R A L T R A N S - V A R * M O R E

• Press SOFT KEY 1 to select CUTOFF Formula.

• Press SOFT KEY 2 to select TRANSFORMATION Formula.
• Press SOFT KEY 3 to select ASSAY VALIDATION Formula.
• Press *MORE to access additional formula types.
• Press SOFT KEY 3 to select TRANS-VAR.
• Press ENTER to save the selected formula type and advance to the next
screen.

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Validation Type Selection

If you selected VAL, this option allows you to choose which Validation
Formula type (Control, Assay, or Blank Validation formulas) to enter for the
assay.

S E L E C T V A L I D A T I O N T Y P E :
C O N T R O L A S S A Y B L A N K

• Press SOFT KEY 1 to select Control Validation Formula.

• Press SOFT KEY 3 to select Assay Validation Formula.
• Press SOFT KEY 4 to select Blank Validation Formula.
• Press ENTER to save the validation type and advance.

Formula Entry

™ Note: In formulas, “OD” is used to represent the optical density

value.

F O R M U L A # 1 :
M A T H O T H E R M A P F U N C T N

• After a moment, the FORMULA #1: prompt disappears, and the

formula can be entered.
• Each formula can contain a maximum of 24 characters. Spaces are
unnecessary.
• Use the LEFT and RIGHT ARROW keys to move the cursor to the
previous or next editable field.
• Press SOFT KEY 1 to place the next item on the MATH list at the cursor
position. The following table shows the order of items on the MATH list:

+ Addition sign

- Subtraction sign

* Multiplication sign

/ Division sign

% Percent

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56 | Chapter 3: Operation

= Equal

> Greater than

>= Greater than or equal to

< Less than

<= Less than or Equal to

™ Note: The reader software checks the formulas for errors during data
reduction. A syntax error in a formula will result in a “Token Error” on
results reports.

• Press SOFT KEY 2 to place the next item on the OTHER list at the
cursor position. See the table that follows for the order of items on the
OTHER list:

( Left parenthesis

) Right parenthesis

AND Logical AND

OR Logical OR

• Press SOFT KEY 3 to place the next defined item on the plate map list
(i.e., STD, NC, PC, BLK) at the cursor position.
• Press SOFT KEY 4 to place the next option on the FUNCTION list at
the cursor position. The available functions are:

LOG10 Log Base 10

ALOG1 Anti Log Base 10

0

AB Absolute Value

PWR Power

ALOG Anti Log

LOG Log

SQRT Square Root

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EXAMPLES:

LOG10: Log Base 10 ALOG: Anti Log

Log10 2 = 0.301029995 ALOG (0.69314718) = 2
ALOG10: Anti Log Base 10 LOG: Log
ALOG10 (0.30102995) = 2 LOG 2 = 0.69314718
AB: Absolute Value SQRT: Square Root
AB (-1) = 1 SQRT 2 = 1.4142
PWR: Power
(10 PWR 2) = 100

• Well locations, well types, or numbers in parentheses precede functions.

• Press the Clear key to clear the item displayed at the cursor position.
• Press the Shift and Clear keys to clear the entire entry.
• Press Enter to save the displayed value and advance to the next screen,
or use the Previous Screen key to move backward through the menu
structure.

Number of Required Controls/Blanks

If a control or blank validation formula is entered, this screen allows you to

enter the number of valid controls / blanks for the assay. Any previously
defined values will appear on the display.

P C : N U M B E R O F V A L I D
R E P L I C A T E S R E Q U I R E D ? 0 2

• Use the Numeric keys to enter the Number of Required Controls. The
range is 1 through the number of defined replicates of a control or
blank.
• Press the Clear key to clear the displayed value.
• Press the Enter key to save the displayed value and advance to the next
screen, or use the Previous Screen key to move backward through the
menu structure.

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58 | Chapter 3: Operation

Cutoff Formulas

A cutoff formula calculates a cutoff value that is used for classifying samples.
During data reduction, results are evaluated against the cutoff value (with an
optional greyzone), and each well is assigned a call POS (positive), NEG
(negative), or EQUIV (equivocal).
• One cutoff formula may be defined per assay.
• If Transformation Formulas are defined, cutoffs are based on the
transformed results. Refer to “FORMULA” on page 53 for the order in
which formulas are processed.
• A cutoff formula can consist of a simple numeric value (1.500), a well
identifier (PC or PC;x to represent the mean), or a formula combining
the two (NC;x+0.050).
• A “greyzone” around the cutoff value can be defined, to indicate
equivocal or indeterminate results.
• Do not use the < or > mathematical symbols in a cutoff formula.
• Tip: Choose to print a Column Report to see the greyzone and cutoff
values as well as the equations used to assign calls to samples.
After selecting an assay (page 32), define the Cutoff formula as shown below:

D E F I N E :
M E T H O D M A P F O R M U L A C U R V E
Ï

S E L E C T F O R M U L A T Y P E :
C U T O F F T R A N S V A L * M O R E
Ï

F O R M U L A # 1 :
M A T H O T H E R M A P F U N C T N

After the FORMULA #1: prompt disappears, enter the formula as described in
“Formula Entry” on page 55. Refer to the examples on page 60.

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Greyzone Entry

The greyzone is a definable area around the cutoff value. Samples falling
within an area defined by the greyzone (ex. ± 5.0% of the cutoff value) could be
considered equivocal (EQUIV).

E N T E R G R E Y Z O N E : 0 5 %

• Use the NUMERIC keys to enter the greyzone percentage.

• The valid entry range is from 00 to 99%. An entry of 00% indicates no
greyzone, although a sample equal to the cutoff value will still receive
the EQUIV call.
• See POS / NEG Calls below for information on how calls are assigned.
• Press the Clear key to clear the displayed value.
• Press the Enter key to save the displayed value and advance.

Positive/Negative Calls for Cutoff

After the greyzone is defined, calls for the sample wells (POSitive, NEGative,
EQUIVocal) must be defined.

S A M P L E > C U T O F F + 0 5 % : P O S
P O S N E G

• Select POS or NEG to select the call that will be assigned to samples
greater than the cutoff value plus the greyzone.
• If, for example, POS is selected as shown in the above screen, calls will
be assigned according to the following equations (SMP represents the
sample wells):

EQUIV: SMP <= (CUTOFF+(CUTOFF*GREYZONE)) AND

SMP >= (CUTOFF-(CUTOFF*GREYZONE))

POS: SMP > (CUTOFF+(CUTOFF*GREYZONE))

NEG: SMP < (CUTOFF-(CUTOFF*GREYZONE))

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60 | Chapter 3: Operation

Examples

1: The cutoff between negative and positive calls should be calculated as the
average of the negative controls plus the OD value of 0.500. Samples greater than
the cutoff should be labeled as positive. No greyzone is required.
• For this example, NC;x (the mean of the NC wells) equals 1.000 OD
• The cutoff formula is NC;x+0.5
• The greyzone is 00%
• POS is selected for SAMP>CUTOFF+00%
• Calls are assigned to sample wells as follows:
¾ EQUIV if the sample equals 1.500
¾ POS if the sample is greater than 1.500
¾ NEG if the sample is less than 1.500
2: For a quantitative assay, samples with OD values greater than the STD2 mean
plus a 10% greyzone should be labeled as positive; samples with OD values less
than the STD2 mean minus the 10% greyzone should be labeled as negative. All
other samples should be considered equivocal.
• For this example, STD2;x (the mean of the STD2 wells) equals 2.000 OD
• The cutoff formula is simply STD2;x
• The greyzone is 10%
• POS is selected for SAMP>CUTOFF+10%
• Calls are assigned to sample wells as follows:

¾ EQUIV if the sample is greater than or equal to 1.800 and

less than or equal to 2.200
¾ POS if the sample is greater than 2.200
¾ NEG if the sample is less than 1.800

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 Defining Assays | 61

Transformation Formulas

Transformation formulas change the absorbance data of all wells defined in the
Map to another format, in preparation for further data reduction.

Transformation Formula Definition

• From the assay Define Menu, press the arrow corresponding to

Formula.

D E F I N E :
M E T H O D M A P F O R M U L A C U R V E

Ï
• This will bring you to a screen asking to Select Formula Type. At this
screen, select TRANS and then enter the formula using the Math,
Other, Map and Function keys.

S E L E C T F O R M U L A T Y P E :
C U T O F F T R A N S V A L * M O R E

Example: Divide all ODs on the plate by 2 and multiply by 100.

Enter the formula: (OD/2)*100

This formula will be applied to the ODs of all samples, standards,

controls, and blanks that are present on the plate map.

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62 | Chapter 3: Operation

Transformation Scope Variable

For more complex transformations, a Transformation Scope Variable

(TVar) can be defined for use with a transformation formula. This variable
defines the scope of the transformation: whether to apply the transformation to
just the samples (SMP) or to all wells defined on the plate (OD).
From the assay Define Menu, press the arrow corresponding to Formula.

D E F I N E :
M E T H O D M A P F O R M U L A C U R V E

Ï
This will bring you to a screen prompting you to Select Formula Type. Press
*MORE at this screen.

S E L E C T F O R M U L A T Y P E :
C U T O F F T R A N S V A L * M O R E

Ï
The options displayed now include TRANS-VAR. Press VAR at this screen.

S E L E C T F O R M U L A T Y P E :
G E N E R A L T R A N S - V A R * M O R E

Ï
The following screen will appear, asking you to choose the scope of this
transformation.

S C O P E V A R I A B L E : O D
S M P O D

If SMP is chosen, the transformation formula will be applied only to the

samples defined in the plate map. If OD is chosen, the formula definition
screen will appear. Use the formula keys (Math, Other, Map and Function) to
define the transformation variable (TVar). Once the variable has been
defined, it can be used in a transformation formula. The TVar will be available
as a MAP option when writing the transformation formula.

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 Defining Assays | 63

Example: An assay plate map has 2 blanks, 1 control well in duplicate (CTL1),
1 negative control well in triplicate (NC), and 5 standards in duplicate
(STD1-STD5) with varying concentrations.
The assay data reduction states:
• Subtract the mean of CTL1 from the mean of the NC. Subtract the
difference from all ODs on the plate.
• Divide the result of the above by the means of the NC less the means of
CTL1, and then multiply by 100.
On paper, the formula reads:
(OD-(NC;x-CTL1;x))/(NC;x-CTL1;x)*100
• On the reader, define the formula (NC;x-CTL1;x) as the Transformation
Variable, since the transformation will apply to all standards, controls
and samples on the plate.
• At the SCOPE VARIABLE screen, choose OD and press Enter. Now
enter the formula (NC;x-CTL1;x) by using the MATH, OTHER, MAP
and FUNCTION keys. Press the ENTER key.
• The formula definition screen is displayed. Choose TRANS.
• Enter the following formula: (OD-(TVar))/(TVar)*100 using the
MATH, OTHER, MAP, and FUNCTION keys. (TVar is included in Map
options on the formula entry screen.) The transformation formula has
been added to the assay definition.

Another Transformation Example:

In the case of competitive reactions, converting absorbance data to percent

B/B0 can be: (OD/Std1)*100. This divides all the wells by STD1 (presumably
the 0 standard), and multiplies the results by 100.
• At the SCOPE VARIABLE screen, choose OD and press Enter.
• Select STD1 from MAP and Press Enter.
• Choose TRANS.
• Enter (OD/TVAR) * 100

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64 | Chapter 3: Operation

CURVE

The CURVE entry screens allow editing and entry of:

• Curve-Fit Type
• Editing of Outliers
• Axis Identification
• Extrapolation of Unknowns
These screens are displayed on the ELx808 in the order in which they appear in the
assay. If a closed variable (i.e., an element of the assay definition that you cannot
access or modify) is being used in the assay, the entry screen is omitted.

Curve Fit

The Curve-Fit screen allows you to select the curve-fit method that will be
applied to the assay. Any previously defined curve-fit type appears on the top
line of the display, and available options on the second line.
The Curve-Fit screen has three sub-menu screens. Each sub-menu screen
provides different curve-fit options for selection. These options include C-
Spline, Linear, Quadratic, Cubic, 4-P, 2-P (Logit/Log), PT to PT (point to
point), and None.
• Linear curve fit: A simple best-fit straight line is plotted using the
values of the standards.
• Quadratic or “Quad” curve fit: A curve fit which uses the Quadratic
equation “ax2 + bx + c = y” to plot the standard's values. Utilizing this
curve, any data point for a standard that deviates from the ideal value
will not affect the entire curve.
• Cubic curve fit: A curve fit that uses the equation “ax3 + bx2 + cx + d =
y” to plot the standard's values. This type of curve fit is affected even
less than the quadratic fit when any particular standard has a poor
value.
• 2-P (LOGIT/LOG): A curve fitted to the standard values, which is
characterized by a skewed sigmoidal (S-shaped) plot that eventually
becomes asymptotic to the upper and lower standard values. The
logistic equation is algebraically transformed to a simpler form in which
experimentally determined values are used for the responses at
concentrations of zero and infinity.
• Cubic Spline (C-Spline) curve fit: A piecewise polynomial
approximation consisting of joining a set of data points by a series of
straight lines, which is then smoothed by using a cubic fit.

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 Defining Assays | 65

• 4-Parameter Logistic or “4-P”: A curve fitted to the standard

values, which is characterized by a skewed sigmoidal (S-shaped) plot
that eventually becomes asymptotic to the upper and lower standard
values. The 4 parameters are: Left asymptote, Right asymptote, Slope
and Value at the Inflection point. This fit is most recommended for
immunoassay data, and is more exact than Logit/Log.
• Point to Point or “PT to PT”: A plot that connects each standard
point with a line, with no averaging of the values to “smooth” the curve
at each standard.

C U R V E - F I T T Y P E : C - S P L I N E
N O N E L I N E A R Q U A D * M O R E

C U R V E - F I T T Y P E : C - S P L I N E
C U B I C 4 - P 2 - P * M O R E

C U R V E - F I T T Y P E : C - S P L I N E
C - S P L I N E P T - P T * M O R E

• Press SOFT KEYS 1, 2, 3, or 4 to select the curve-fit type that is

displayed above the soft key. Select the soft key below the menu option
MORE to display additional options. The top line of the display updates
to reflect this selection.
• Press ENTER to save the selection and advance to the next screen, or
use the MAIN MENU and PREVIOUS SCREEN keys to move
backward through the menu structure.

Edit Standard Outliers

This screen allows you to select which method (None or Manual) will be used
to edit Standard Outlier values. After the standard curve has been calculated,
one or more standards can be excluded from the recalculation of the curve.
Any previously defined edit method is displayed.

E D I T S T D O U T L I E R S : M A N U A L
N O N E M A N U A L

• Press SOFT KEY 1 or 2 to select the edit option displayed above the
soft key. The display updates to reflect your selection.
• Select NONE to suppress the Edit Standard Outliers capability for this
assay.

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66 | Chapter 3: Operation

• Choose MANUAL to enable the capability.

¾ If AVERAGE STANDARDS is set to NO, the individual standard
replicates are available for editing. If set to YES, the standard groups
are available for editing.
¾ After the assay is run and reports are generated, press REPORT
from the Main Menu. Press RESULT, select the assay, and then
press ENTER. The EDIT STD OUTLIERS? YES/NO prompt will
appear. See Editing Standard Outliers on page 74 for further
instructions.
• Use the ENTER key to save the selection and advance to the next
screen, or use the MAIN MENU and PREVIOUS SCREEN keys to move
backward through the menu structure.

Axis Selection

This screen allows you to select the X and Y Axis Type. Any previously defined
axis type will be displayed. This option screen appears only if Manual Map
Generation has been selected.

X / Y A X I S T Y P E : L I N
L I N L I N / L O G L O G L O G / L I N

• Press SOFT KEY 1, 2, 3, or 4 to select the method by which the X and

Y-axes will be scaled. The top line of the display updates to reflect the
selection.
• This option is not available for the 2-P and 4-P curve-fit types. The X/Y
scaling for these curves is always LIN/LIN.
• Press ENTER to save the selection and advance to the next screen.

Extrapolation of Unknowns

This screen allows you to choose whether to extrapolate the curve to evaluate
samples outside of the absorbance range defined by the standards. Any
previously defined decision appears on the screen.

E X T R A P O L A T E U N K N O W N S ? Y E S
Y E S N O

• Press SOFT KEY 1 to select YES (extrapolate the unknowns). The top
line of the display updates to reflect this selection.
• On the printed reports, extrapolated concentrations (RSLT values) are
surrounded by < > (e.g., <44.425>).

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 Defining Assays | 67

• Press SOFT KEY 2 to select NO. The top line of the display updates to
reflect this selection.

™ Note: If extrapolation is chosen for the Point-to-Point curve fit, unknown

concentrations will be extrapolated linearly from the nearest segment of
the curve. If the plot includes both increasing and decreasing segments,
the curve printout will be labeled “Ambiguous.” The resulting values,
which actually are extrapolated, may not be indicated as such. All
calculated results for an “Ambiguous” curve should be considered
unreliable.

Panel Assays

A Panel assay is a collection of up to 8 assays to be run on one plate.

• A common reason to use a Panel assay is when one or more samples are
tested for more than one antigen. An example is an ENA panel, which
could screen dsDNA, Sm, SSA, SSB, Scl-70, and/or Jo-1 on one microplate.
• Only one Panel assay can be defined on the reader at any time.
• The assays specified within the Panel must be predefined in any of the
assay positions 1-55.
• The assays specified within the Panel must all use the Endpoint read
method.
• The assays specified within the Panel must all read at the same
wavelength(s).
• Any curve-fit type, formulas, or standard concentrations previously
defined for each assay will be used when the assay is selected for a Panel.
• Panel assays cannot reuse standard curves.
• Only an Auto map is recognized by a Panel assay. Custom locations
entered using a Manual map will be overwritten by the Panel assay.
• If the Panel runs in a 1*12(ACROSS) configuration, both the Map Direction
and Replication Direction must be set to ACROSS during assay definition.
ACROSS would also then be selected as the direction during Panel
definition.
• The type and number of controls, blanks, standards, and replicates in the
assays chosen for the Panel will be “copied” into the Panel definition. Map
or assay parameters must first be changed in the predefined assay before
they can change in the Panel.
To create a panel assay, start at the Main Menu, select DEFINE, then choose assay
number 99. Enter the panel assay name.

N A M E : P A N E L

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68 | Chapter 3: Operation

- / : S P A C E

• The default name is “PANEL”.

• Use the alpha and numeric keys to update the assay name, if desired.
• Press Enter to continue. The Number of Assays entry screen will appear.

N U M B E R O F A S S A Y S : 2

• Specify the number of assays to include in the panel (1 to 8).

• Press Enter to continue. The Mapping Direction selection screen will
appear.

M A P P I N G D I R E C T I O N : D O W N
D O W N A C R O S S

• This option ensures that all assays will be mapped in the same direction.
• Select DOWN or ACROSS.
After selecting the mapping direction of the assays, choose which assays to
include in the panel.

S E L E C T A S S A Y N U M B E R : 2 2
N A M E : H B S - A G 1

• Press the Options key to cycle through the assay numbers and names, or
use the numeric keys to enter an assay number. Press Enter to make a
selection.
• After an assay is selected, its starting location must be defined.

S T A R T M A P P I N G
A T W E L L L O C A T I O N : A 0 1

• Use the alpha and numeric keys to choose the well location to begin the
assay.
• Repeat this process for each assay within the panel. Remain aware of the
total number of controls, standards, and blanks that were originally
mapped in each individual assay while mapping for the Panel assay.

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 Defining Assays | 69

• For example, to include Assays 1, 8, and 22 in the Panel assay (DOWN

mapping is selected for the Panel):

¾ Assay 1 has a total of 12 wells defined for controls, blanks, and

standards. In the Panel, the mapping for Assay 1 begins in well A01. The
user wants to run 6 samples in Assay 1. Assay 1 now fills wells A01
through B03.
¾ The mapping for Assay 8 can begin in well B04, or any well other than
A01 to B03. The reader will “beep” if you try to map into a well that is
already assigned for use with the Panel.
¾ The mapping for Assay 22 may begin at the next available well location
after Assay 8 mapping is complete.
¾ The Panel Assay results are sorted by Sample (unless a custom assay has
been programmed by BioTek).

™ Note: The Interpretation of Results reports for each assay in

the Panel will print first, and then the Sample results will print.

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70 | Chapter 3: Operation

Reading a Microplate
Press the READ option, found at the Main Menu, to read a microplate.
• From the MAIN MENU screen, press the soft key beneath the READ menu
option to access the SELECT ASSAY NUMBER screen.
• Alternately, press the red READ key on the lower right of the keyboard.

Select Assay

At the Select Assay Number screens:

• Use the NUMERIC keys to enter the number of any predefined Assay
Definition Files stored in the reader’s memory, or the OPTION key to
advance one assay at a time. The cursor is positioned at the first editable
field and advances automatically. The numeric range depends on the
number of assays (1-55) programmed in the reader’s memory.
The assay’s name and number are displayed on the screen.

S E L E C T A S S A Y N U M B E R : 2 5
N A M E : H B S - A G

• Press Enter to advance to the Run-Time prompts.

• MAIN MENU: Returns the display to the Main Menu screen.

Run-Time Prompts

After the assay is selected, one or more informational prompts may be presented,
depending on preferences selected in UTIL > READ, whether or not the assay
specifies manual mapping, or if a custom assay database is installed on the reader.
• Prompts enabled via UTIL > READ can include ENTER NUMBER OF
SAMPLES, PLATE ID, and ENTER SAMPLE ID.

• If the assay specifies manual mapping, prompts for information will

include the locations for the sample wells.
• If running a custom assay, typical prompts might include:
Number of samples
Standard concentrations
Assay ID
Fill pattern
Blank method
First well location
Replicate count for each well type

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 Reading a Microplate | 71

Wavelength mode
Report preferences

Enter Number of Samples

Use this screen to enter from 00 to the maximum number of samples permitted by
the previously created well map. If there are no controls, standards, or blanks
defined, the minimum number of samples is 1.
This value controls the number of samples reported if Matrix or Column reports
are requested.

E N T E R N U M B E R O F
S A M P L E S : 9 1

Enter Plate ID

You can enter a 10-character (maximum) identifier to assign to the plate. Since this
Plate ID will be stored in the reader’s memory, each plate ID should be unique.

™ Note: Use caution when creating multiple Plate IDs. The reader does
not warn you that you are about to exceed the maximum of 10 plate
IDs stored in memory. If an eleventh Plate ID is added, it will
overwrite the first Plate ID stored in memory.

™ Note: If the internal bar code scanner option is installed, the reader
will automatically scan the plate/bar code label and use this as the
Plate ID.

P L A T E I D :
- / : S P A C E

• Use the KEYPAD to enter numbers, and the LEFT / RIGHT arrow keys to
move the cursor. Clear clears the display.

Enter Sample ID

You can start entering a starting sample identification number from 0001 to 9999.
The software will automatically increment each subsequent sample identification
number by 1. Sample IDs will be assigned according to the previously defined
mapping order.

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72 | Chapter 3: Operation

E N T E R
S A M P L E I D : 0 0 0 1

• Use the KEYPAD to enter numbers and the LEFT / RIGHT arrow keys to
move the cursor. Clear clears the display.

Prompts for Well Location

Well locations can be changed at run time if a Manual Map has been specified, and
you have requested a sample count at run time via the Utilities menu.

S A M P L E # 1 L O C A T I O N
R E P # 1 : A 0 1

• Use the KEYBOARD to enter the well location, using the SHIFT-LETTER
sequence to key in letters, and press Enter to specify the desired location.

Beginning the Plate Read

When the following screen appears on the display, the reader is ready to read a
plate:

P L A C E P L A T E I N C A R R I E R
A N D P R E S S < R E A D > K E Y

• Place the plate in the carrier and press the READ key to initiate the plate
read. After the plate has been read, all requested reports will be generated.
• To halt in read in progress, press the STOP key.

™ Note: If using the incubation option, the reader will wait for the
incubator to reach temperature before reading the plate.

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 Printing Reports and Assay Lists | 73

Printing Reports and Assay Lists

Important! The 'OUT' indication on reports means the OD

for an individual well, or the average OD for a group of
wells, falls outside the minimum/maximum OD range
defined for the assay. For the Quick Read assay on the
ELx808, this range is -3.0 OD to +3.0 OD. For assays defined
onboard by a user (Assay02-Assay55), it is -4.0 OD to +4.0
OD.

Reports are automatically generated after a plate has been read (see OUTPUT Options
on page 19 and REPORT Type on page 19 in Chapter 2 for information on selecting
reports). To manually regenerate results reports, use the REPORT option from the
Main Menu. You can also print Map, Assay, and Assay List reports.

™ See Appendix A for sample reports.

R E A D Y 9 : 4 5 P M 0 5 / 0 9 / 0 7
R E A D D E F I N E R E P O R T U T I L

P R I N T R E P O R T :
R E S U L T M A P A S S A Y L I S T

• The eight most recent sets of plate data are stored in memory (see next page).
Select REPORT > RESULT to print an exact copy of results from the plate
reading.

¾ The form in which the results are presented is determined by the report
settings (Matrix, Column, Curve Fit) specified under UTIL > OUTPUT.
• Select MAP to print a matrix showing the locations of the Blanks, Standards,
Controls, and Samples for a selected assay.
• Select ASSAY to print a plate map and a listing of all of the assay’s settings,
such as wavelengths, numbers of well types, formulas, and curve-fit
parameters.
• Select LIST to print a list of all assays (name and number) currently
programmed in the ELx808.

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74 | Chapter 3: Operation

™ Note: The reader stores measurement values for the last 8 plates in
memory.

Result

R E P O R T : H B S - A G
I D : 0 0 1 0 7 / 1 7 / 0 7

• Use the OPTION key to select the appropriate Plate ID and Report. Note
that the Assay ID will change if the selected Plate ID was read with a
different assay. Once you have found the correct Plate ID, press Enter.

Editing Standard Outliers

If a standard curve was generated and if EDIT STANDARD OUTLIERS was set to
MANUAL in the assay definition, the option to edit outliers is presented.

E D I T S T D O U T L I E R S :
Y E S N O

• Select NO to include all standards in the curve-fit calculations.

• Select YES to indicate that one or more standard replicates or groups
should be temporarily excluded from curve fit-calculations.

¾ If AVERAGE STANDARDS was set to NO in the assay definition, one or

more standard replicates can be chosen for exclusion.

E D I T S T D 1 R E P 1 ? Y E S
Y E S N O

• Select YES to exclude the replicate from curve-fit calculations.

• Select NO to retain the replicate.
• Press ENTER to advance to the next replicate.

¾ If AVERAGE STANDARDS was set to YES in the assay definition, one

or more standard groups can be chosen for exclusion.

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 Printing Reports and Assay Lists | 75

E D I T S T D 1 ; X ? Y E S
Y E S N O

• Select YES to exclude the group from curve-fit calculations.

• Select NO to retain the group.
• Press ENTER to advance to the next group.

™ Note: Each curve-fit type requires a minimum number of standards for

curve generation: 4 for 2-P, 4-P, cubic, and cubic-spline, 3 for
quadratic, and 2 for linear and point-to-point. Exercise caution when
editing outliers. If the assay is left with insufficient standards, the
curve fit will fail.

Printing Results

After the assay is selected and standard outliers are edited (if necessary), the
results report can be printed.

P R I N T R E S U L T S ?
Y E S N O

• Ensure that the printer is connected, turned on, and filled with paper.
• Press YES to print reports, or NO to return to the Main Menu.

Map

• Select REPORT at the Main Menu, and then select MAP.

S E L E C T A S S A Y N U M B E R : 0 1
N A M E : H B S - A G

• Press Options to cycle through the list of available assays, or enter the
number of the desired assay.
• Press Enter to print the report.

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76 | Chapter 3: Operation

Assay

• Select REPORT at the Main Menu, and then select ASSAY.

S E L E C T A S S A Y N U M B E R : 0 1
N A M E : H B S - A G

• Press Options to cycle through the list of available assays, or enter the
number of the desired assay.
• Press Enter to print the report.

List

• Select REPORT at the Main Menu, and then select LIST. The entire list of
assays stored in the ELx808’s memory will be sent to the printer.

Recommendations for Optimum Performance

• Microplates should be perfectly clean and free of dust or bottom scratches. Use
new microplates from sealed packages.
• Do not allow dust to settle on the surface of the solution; use microplate covers
when not reading the plate. Filter solutions to remove particulates that could cause
erroneous readings.
• Although the ELx808 supports standard flat, U-bottom, and V bottom 96-well
microplates, the reader achieves optimum performance with optically clear, flat-
bottomed wells.
• Non-uniformity in the optical density of the well bottoms can cause loss of
accuracy, especially with U- and V-bottom polyvinyl microplates. Check for this
by reading an empty microplate. Dual wavelength readings can eliminate this
problem, or bring the variation in density readings within acceptable limits for
most measurements.
• Inaccuracy in pipetting has a large effect on measurements, especially if smaller
volumes of liquid are used. For best results, use at least 100 µL per well.
• The inclination of the meniscus can cause loss of accuracy in some solutions,
especially with small volumes. Agitate the microplate before reading to help bring
this problem within acceptable limits. Use Tween 20, if possible (or some other
wetting agent) to normalize the meniscus. Some solutions develop menisci over a
period of several minutes. This effect varies with the brand of microplate and the

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 Recommendations for Optimum Performance | 77

solution composition. As the center of the meniscus drops and shortens the light
path, the density readings change. The meniscus shape will stabilize over time.
• Although the effect of ambient light is mathematically quantified and subtracted
from each absorbance reading, the illumination of the instrument by strong
ambient light should be avoided. If interference from ambient light is suspected,
read a microplate of high-density solutions under the suspect conditions, and
again with all ambient light blocked (in a dark room for example), then compare
results. The blocked results will appear as an increase in optical density readings if
light is influencing the readings. Because of the mathematical correction, this
difference under normal conditions should be slight or nonexistent.
• A 10-minute warm-up of the instrument is suggested, prior to reading, to achieve
the best repeatability from microplate-to-microplate measurements.

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78 | Chapter 3: Operation

BioTek Instruments, Inc.

Chapter 4

Instrument Qualification

This chapter discusses the tasks and procedures necessary for

qualifying instrument performance initially and on an ongoing basis.

Overview ........................................................................ 80 

Recommended Qualification Schedule ................................. 80 
Test Descriptions ............................................................. 81 
System Test ............................................................... 81 
Checksum Test ........................................................... 84 
Absorbance Plate Test .................................................. 85 
Empty Carrier Test ...................................................... 88 
Liquid Tests ................................................................ 88 
Perform the Tests ............................................................ 89 
System Test ............................................................... 89 
Checksum Test ........................................................... 89 
Absorbance Plate Test .................................................. 89 
Empty Carrier Test ...................................................... 91 
Liquid Tests ................................................................ 92 
80 | Chapter 4: Instrument Qualification

Overview
This chapter contains BioTek Instruments’ recommended Installation Qualification (IQ),
Operational Qualification (OQ), and Performance Qualification (PQ) procedures for all
models of the ELx808 Absorbance Microplate Reader.
Every ELx808 reader is fully tested at BioTek prior to shipment and should operate
properly upon initial setup. If you suspect that a problem occurred during shipment, if
you have received the equipment after returning it to the factory for service, and/or if
regulatory requirements dictate that you qualify the equipment on a routine basis, you
should perform the procedures outlined in this chapter.

™ An ELx808 Product Qualification Package (PN 7340543) is available for

purchase. It contains procedures for performing IQ/OQ/PQ and
Preventive Maintenance, with spreadsheets for analyzing results and
checklists and logbooks. Contact your BioTek dealer for more information.

Recommended Qualification Schedule

The schedule below defines the factory-recommended intervals for qualifying an ELx808
reader used two to five days a week. The actual frequency, however, may be adjusted
depending on your usage of the instrument. This schedule assumes the reader is properly
maintained as outlined in the Preventive Maintenance chapter.

™ The risk factors associated with your tests may require that the
Operational and Performance Qualification procedures be performed
more or less frequently than shown here.

Installation Operational Performance

Qualification Qualification Qualification
Tests
Initially/
Initially Monthly Quarterly
Annually
System Self-Test, p. 89 9 9 9
Checksum Test, p. 89 9 9 9
Absorbance Plate Test, p. 89 9 9 9
Empty Carrier Test, p. 91 9 9
Liquid Tests, p. 92
Liquid Test 1*, or
Liquid Test 2* 9 9
Liquid Test 3**, (optional,
for 340 nm)

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 Test Descriptions | 81

* If you have an Absorbance Test Plate, run Liquid Test 1. If you do not have a Test Plate,
run Liquid Test 2.
** Liquid Test 3 is optional; it is provided for sites requiring verification at wavelengths
lower than those attainable with the Absorbance Test Plate.

Test Descriptions

System Test

The System Test runs automatically whenever the instrument is turned on. It can also
be performed manually using Gen5 software, or the keypad (UTIL > TESTS >
SYSTEM). If the power-up System Test fails, the reader will “beep” repeatedly and
display an error code; see Chapter 6.
When run manually, a System Test report is printed (or displayed in Gen5). The report
(sample shown on the next page) includes results for the System Optics Test and
Autocal Analysis. We recommend saving System Test reports to document periodic
testing and assist with troubleshooting.
• The System Test conducts a series of tests at each wavelength defined in the
filter table to confirm adequate light levels, low electronic noise, adequate
photodiode sensitivity, overall system cleanliness, and (if equipped) proper
function of the incubator. The testing is designed to verify that the ELx808 will
give in-specification performance for each set wavelength over the specified
OD range.
• Autocal is run only at the factory (or by an authorized service engineer) to
calibrate the carrier axis of the instrument. The results of this test are included
in the System Test Report. In the field, the alignment of the Absorbance Plate
Test is used to verify that the carrier positioning is correct.

Terms used on the System Test Report

Air Blank A full light reading through a filter with no plate in the light path.
Dark Current A reading taken with the light blocked to measure background
light levels in the reading chamber. Also used as a measure of
background electronic noise within the measurement circuit.
Gain An automatic electronic adjustment to the measurement circuit.
The gain adjustment compensates for changing light levels or
filter variations. For example, if the lamp output decreases
slightly, the gain will increase to make up the difference.
Axis Refers to a motor for the filter wheel or plate carrier.
Offset A numerical limit, usually a range. For example, if the gain fails
an offset test, it may be too high or too low.

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Gen5 System Test Report

Reader: ELx808 (Serial Number: 118943)

Basecode: P/N 7340201 (v3.15)
Date and Time: 7/28/2012 3:06:11 PM
User: Administrator
Company: BioTek
Comments: System Test run during the IQ

Test Results

Operator ID:______________________________________________________________

Notes:____________________________________________________________________

03:11PM 7/28/12 SYSTEM SELF TEST

Filter: 405 Gain: 15.06

Channel: Ref 1 2 3 4 5 6 7 8
Air: 24272 36673 54326 49036 37302 41392 37956 46880 42789
Dark: 1627 1790 1738 1983 1922 1969 2155 1978 2041
Delta: 22645 34883 52588 47053 35380 39423 35801 44902 40748

Filter: 450 Gain: 5.12

Channel: Ref 1 2 3 4 5 6 7 8
Air: 26520 40514 54985 51166 41113 45699 43595 50055 47401
Dark: 587 644 626 708 688 705 767 707 729
Delta: 25933 39870 54359 50458 40425 44994 42828 49348 46672

Filter: 490 Gain: 16.00

Channel: Ref 1 2 3 4 5 6 7 8
Air: 29585 40785 53183 50389 42324 46927 45225 51112 48181
Dark: 1806 1981 1927 2187 2121 2172 2369 2182 2247
Delta: 27779 38804 51256 48202 40203 44755 42856 48930 45934

Filter: 620 Gain: 8.26

Channel: Ref 1 2 3 4 5 6 7 8
Air: 28973 40210 53315 49573 42596 47522 46312 50677 49083
Dark: 922 1012 983 1116 1084 1111 1211 1114 1150
Delta: 28051 39198 52332 48457 41512 46411 45101 49563 47933

Filter: 630 Gain: 5.57

Channel: Ref 1 2 3 4 5 6 7 8
Air: 28127 42576 54097 52142 44139 48634 47638 52007 50305
Dark: 630 691 672 762 739 758 826 760 784
Delta: 27497 41885 53425 51380 43400 47876 46812 51247 49521

Filter: 340 Gain: 64.00

Channel: Ref 1 2 3 4 5 6 7 8
Air: 18961 28041 46573 41370 27548 30291 25144 36515 31946
Dark: 7163 7855 7642 8677 8411 8630 9413 8655 8927
Delta: 11798 20186 38931 32693 19137 21661 15731 27860 23019
Channel: Ref 1 2 3 4 5 6 7 8
Noise Max: 3952 4351 4225 4817 4668 4790 5237 4807 4963
Noise Min: 3951 4346 4220 4814 4664 4785 5233 4802 4957
Delta: 1 5 5 3 4 5 4 5 6

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 Test Descriptions | 83

03:11PM 7/28/12 INCUBATOR SELF TEST

Temperature Setpoint: 37.0 Current Average: 36.9 A/D Test: PASS

Zone 1: 37.0 Min: 36.9 Max: 37.0 Range: PASS Thermistor: PASS
Zone 2: 37.0 Min: 37.0 Max: 37.1 Range: PASS Thermistor: PASS
Zone 3: 37.0 Min: 36.9 Max: 37.0 Range: PASS Thermistor: PASS
Zone 4: 36.9 Min: 36.9 Max: 37.0 Range: PASS Thermistor: PASS

The INCUBATOR SELF TEST shows the

PASS or FAIL condition of the four heating
AUTOCAL ANALYSIS zones in the incubator (if equipped). See
Incubator Self-Test on the next page.

Upper Left Corner: x=5344 y=0

Lower Left Corner: x=6156 y=0
Lower Right Corner: x=14850 y=0 AUTOCAL ANALYSIS provides coordinates of
Upper Right Corner: x=14032 y=0 the last Autocal performed on the instrument.
Delta 1 : 5344-6156=-812 AUTOCAL verification is not included in the
Delta 2 : 14032-14850=-818 SYSTEM TEST result given below.

SYSTEM TEST PASS

0000
The SYSTEM TEST result will be either PASS or
FAIL. The SYSTEM TEST result refers only to the
System Optics Test results.

Reviewed/Approved By: ________________________________ Date: ________________

For Technical Support

In the U.S.: In Europe:

BioTek Instruments, Inc. BioTek Instruments GmbH
Tel: 800 242 4685 Tel: 49 (0) 7136-9680
Fax: 802 655 3399 Fax: 49 (0) 7136-968-111

All Others:
Tel: 802 655 4040
Fax: 802 655 3399

email: TAC@biotek.com
Product support center: www.biotek.com/service

Figure 5: Sample System Test Report (it varies slightly depending on how it was run)

™ If a System Test shows one or more channels that have air readings
falling below 30,000 for filter wavelengths of 405 nm or higher, Liquid
Test 1 should be performed. This test is applicable to the eight reading
channels only, not the reference channel.

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Incubator Self Test

The Incubator Self Test of the ELx808 reader is designed to check for hardware
failures (such as open or shorted thermistors, A/D noise) and to verify that each of
the four zones in the incubation system are within ± 2° of the setpoint.
• Temperature Setpoint: The desired temperature programmed by the
user, either through the reader’s keypad or Gen5.
• Current Average: Average temperature of the four zones of the incubator
at the time the system test is run. This number is displayed on the ELx808
front panel and sent to Gen5 if the reader is being computer controlled.
• A/D Test: Checks for allowable noise levels.

03:11PM 7/28/12 INCUBATOR SELF TEST

Temperature Setpoint: 37.0 Current Average: 36.9 A/D Test: PASS

Zone 1: 37.0 Min: 36.9 Max: 37.0 Range: PASS Thermistor: PASS
Zone 2: 37.0 Min: 37.0 Max: 37.1 Range: PASS Thermistor: PASS
Zone 3: 37.0 Min: 36.9 Max: 37.0 Range: PASS Thermistor: PASS
Zone 4: 36.9 Min: 36.9 Max: 37.0 Range: PASS Thermistor: PASS

Zone #: Reports the average temperature of the zone at the time the test is
run. Each zone is evaluated independently.
Min: The minimum temperature recorded during the time that the system test
is run.
Max: Maximum temperature recorded during the time that the system test is
run. There is no range checking between Min and Max temperatures.
Range: This test checks that each zone is within ± 2° of the setpoint.
Thermistor: This test indicates whether the thermistors are functioning or
have failed.

Checksum Test

The Checksum Test compares the basecode software to internally recorded checksum
values to ensure that the programming has not been corrupted. The test runs
automatically when the instrument is turned on, and it can be run manually using the
keypad (UTIL > TESTS > CHKSUM). If the test fails, the reader will “beep”
repeatedly and display an error code; see Chapter 6.
When run manually, the software displays the part numbers and versions of software
currently loaded on the reader. For example:
Software P/N Software Version
7340201 Version 3.21
Code Checksum: (5E35)

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 Test Descriptions | 85

The second screen shows:

Configuration P/N Configuration Version
7340203 Version 3.00

Absorbance Plate Test

This test uses BioTek’s Absorbance Test Plate to confirm the Mechanical Alignment,
Accuracy and Linearity, Repeatability, and Channel-to-Channel uniformity of the
ELx808. The Absorbance Plate Test compares the reader’s optical density
measurements and mechanical alignment to NIST-traceable values. To run the test,
you will need a BioTek Absorbance Test Plate with its accompanying data sheet.

™ The BioTek PN 7260522 Test Plate has a glass filter in location C6, which
is used to check the wavelength accuracy of a monochromator. Although
the ELx808 does not have a monochromator, you must enter values for
the peak wavelength when defining the 7-filter test plate in Gen5
software. Refer to the software help system for more information.

A sample Absorbance Plate Test report is shown on the next page.

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Figure 6: Sample Absorbance Plate Test Report

™ The report varies slightly depending on how it was run (via the keypad or
Gen5).

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 Test Descriptions | 87

™ Important! The Absorbance Plate Test tests the accuracy and

repeatability specifications from 0.000 to 2.500 OD only. In the example
above, the Analysis report displays the OD values read in well positions D4
and E9 but does not indicate PASS or FAIL, because the values are higher
than 2.500 OD and therefore are not within the test range.

If any of the test parameters report as FAIL, confirm that the standard values on the
test plate data sheet match the values on the printout. If there are any mismatches,
correct them and re-run the test. See also the troubleshooting tips below. If the test
continues to fail, contact BioTek TAC. Please have a copy of the test and the reader’s
serial number ready.
• Mechanical Alignment: The Test Plate has several groups of precisely
machined holes to confirm the mechanical alignment of different
microplate readers. The amount of light that shines through these holes is
an indication of how well the reader is aligned. A reading of more than
0.015 OD for any of the designated alignment holes indicates that the light
is being “clipped” and the reader may be out of alignment. If the test fails:
¾ Examine the microplate carrier to ensure that it is clear of debris.
¾ Make sure the Test Plate is properly seated in the microplate carrier.
¾ Check the Test Plate’s alignment holes to ensure they are clear of
debris.
• Accuracy/Linearity: The Test Plate contains neutral-density glass filters
of known OD values at several wavelengths. Actual measurements are
compared against the expected values provided in the Test Plate’s data
sheet. Since there are several filters with differing OD values, the accuracy
across a range of ODs can be established. Once it is proven that the reader
is accurate at these OD values, the reader is also considered to be linear. If
the test fails:
¾ Check the filters on the test plate to ensure they are clean. If
necessary, clean them with lens paper. Do not remove the filters from
the test plate, and do not use alcohol or other cleaning agents.
¾ Ensure that the filter calibration values entered are the same as those
on the Test Plate data sheet.
¾ Ensure that the Test Plate is within its calibration certification period.
• Repeatability: This test ensures the reader meets its repeatability
specification by reading each neutral-density filter on the Test Plate twice
with the filter in the same location. Note that there may not be a Pass/Fail
indication for filter values that are beyond the specified accuracy (and thus
repeatability) range of the instrument. If the test fails:
¾ Examine the glass filters on the test plate to see if there are any loose
particles that may have shifted between readings and caused changes.
If needed, clean the filters with lens paper.

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™ Do not remove the filters from the test plate, and do not
use alcohol or other cleaning agents to clean them.

¾ Check the microplate carrier to ensure that it is clear of debris.

• Channel-to-Channel Variation: This test ensures that selected channels
read the same value for a filter as their paired channel when the plate is
“turned around” 180° in the plate carrier. It is a way to verify that each
channel’s Delta values are adjusted relative to the signal strength of the
reference channel.

Empty Carrier Test

The Empty Carrier Test confirms the ELx808’s read capabilities at the 100% light level,
and can help to identify deteriorating interference wavelength filters and other optical
problems.

Liquid Tests

Conducting Liquid Tests confirms the ELx808’s ability to perform to specification with
liquid samples. Liquid testing differs from testing with the Absorbance Test Plate in
that liquid in the wells has a meniscus, while the Test Plate’s neutral density filters do
not. The optics characteristics may differ in these two cases, alerting you to different
types of problems. The Absorbance Test Plate will indicate the absolute amount of light
absorbed, which will effectively test the linearity of the electronics. The liquid tests will
help detect optical defects such as dirt on the lenses or other contamination that can
contribute to errant readings.
• Liquid Test 1 tests the alignment, accuracy, repeatability, and channel-to-
channel variability of the reader, making evident any problems with the
optics of the system. If you have the Absorbance Test Plate, you will only
need to run Liquid Test 1 for routine testing.
• If you do not have the Absorbance Test Plate, test the linearity,
repeatability, and alignment of the reader by preparing a series of solutions
of varying absorbances as described in Liquid Test 2.
• Liquid Test 3 is an optional test at 340 nm, offered for sites that must have
proof of linearity at wavelengths lower than those attainable with the
Absorbance Test Plate. This test is considered optional because the reader
has good “front end” linearity throughout its wavelength range.

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Perform the Tests

System Test

™ If the System Test fails, see Chapter 6 for troubleshooting tips and
error code descriptions.

Using Gen5, select the following in the appropriate software:

• Open the system menu and select Diagnostics > Run System Test
Using the instrument keypad:
1. Attach a printer to the instrument.
2. From the Main Menu, select UTIL > TESTS > SYSTEM.

Checksum Test

There is no Checksum Test in Gen5. To obtain reader software information:

• Open the system menu and select Reader Control > ELx808. To obtain
Gen5 software information, select Help > About Gen5.
Using the instrument keypad:
• From the main menu, select UTIL > TESTS > CHKSUM.
¾ The initial checksum test display will show the onboard (base code)
software part number, version number, and checksum.
¾ After a few moments, a second screen will show the assay configuration
software part number and version number.

Absorbance Plate Test

Define the Test Plate Parameters

Using Gen5 (click Help for additional instructions):

• Open the system menu and select Diagnostics > Test Plates >
Add/Modify Plates, and click Add.
Using the keypad:
1. From the main menu select UTIL > SETUP > *MORE > CALPLATE. The
Calibration Filter screen will appear.
2. Select a filter wavelength and press Enter. The Wavelength / Calibration
Value screen will appear.

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3. Enter the values listed on the Test Plate data sheet. After each entry, press
Enter to advance to the next well location.
4. Repeat for the remaining filters.
5. When all values have been entered, press the Main Menu key.

Run the Plate Test

™ Before running the test, ensure that the reader is not running in Rapid
Mode. To check this, select UTIL > READ and then cycle through the
prompts until READ IN RAPID MODE? is displayed. Choose NO for
an accurate result.

Using Gen5 (click Help for additional instructions):

• Open the system menu and select Diagnostics > Test Plates > Run.
Using the keypad:
1. Select Start at the Main Menu and select UTIL > TESTS > CALPLATE. The
Calibration Filter screen will appear.
2. Select the appropriate wavelength and press Enter.
3. When prompted, insert the Test Plate into the plate carrier, and press the
READ key to begin the test. The Test Plate Analysis Report (sample on page
86) will be sent to a printer when the test is complete.

Gen5 version 1.07 and earlier users: Gen5’s Reader

Diagnostics Utility (PN 5320201) must be installed before the
Absorbance Plate Test can be performed using Gen5.

Important! The Absorbance Plate Test and CALPLATE

utility in the reader are used only to verify the reader’s
performance. The Test Plate and utility do not correct or
calibrate the instrument.

The Absorbance Plate Test tests the accuracy and repeatability

of the reader to 2.500 OD. It does not indicate a PASS or FAIL
above 2.500 OD.

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Empty Carrier Test

Perform these steps (do not place a microplate on the carrier):

1. For each filter in the filter wheel, run a simple endpoint protocol and then
print the results. (Do not apply blanking or transformations.)
2. Examine the OD values on the printouts. Every OD should read 0.000
± 0.004 OD.
™ These limits are required by BioTek and cannot be changed.

3. Sign and date the printouts, and store them with your test documentation.

If the Empty Carrier test fails at more than one wavelength:

• Check the microplate carrier to ensure that it is clear of debris.
• Verify that the lamp is properly aligned, and then run the test again. If the
lamp has been in use for approximately 600 hours, it may need to be
replaced, and the contacts in the lamp socket may need to be cleaned. Refer
to Replace the Lamp and Clean the Contacts in Chapter 5.
• Follow the steps in Inspect and Clean the Wavelength Filters in
Chapter 5 to remove the filter wheel and clean the filters.

If the Empty Carrier test fails at just one wavelength:

• Remove the filter wheel and examine the filter that exhibited the problem;
it may need to be replaced (contact BioTek). Check for spotting or a halo
effect.

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Liquid Tests

Prepare Test Solutions

BioTek offers a dye solution (PN 7120779, 25 mL; or 7120782, 125 mL) that may be
used in the stock solution formulation for Liquid Tests 1 and 2, or, if you prefer,
you may use the dye solution described below. The purpose of the formulation is
to create a solution that absorbs light at ~2.000 OD full strength when dispensed at
200 µL/well in a flat-bottom microplate.
Alternatively, any solution that gives a stable color will suffice. (This includes
substrates incubated with an enzyme preparation and then stopped with an acidic
or basic solution.) Some enzyme/substrate combinations that may be used as
alternates to the described dye are shown below:
Typical Enzyme-Substrate Combinations and Stopping Solutions

Enzyme Substrate Stopping Solution

Alkaline Phosphate o-nitrophenyl phosphate 3N sodium hydroxide

beta-Galactosidase o-nitrophenyl -beta-D 1M sodium carbonate

galactopyranoside
Peroxidase 2,2'-Azino di-ethylbenzothiazoline- citrate-phosphate buffer,
sulfonic acid (ABTS) pH 2.8
Peroxidase o-phenylenediamine 0.03N sulfuric acid

Stock Solution Formulation

The stock solution for Liquid Tests 1 and 2 may be formulated using the materials
listed below (Solution A), or by diluting a dye solution available from BioTek
(Solution B).

™ Either solution should result in an absorbance of about 2.000 when

using 200 µL in a flat-bottom microwell at 405 nm. The OD value will be
proportional to the volume in the well and the amount of FD&C No. 5
dye or QC Check Solution used. You can use a larger or smaller well
volume, or add more dye or water to adjust the solution. Note that too
small a well volume may result in increased pipetting-related errors.

Solution A

Required Materials:
• BioTek QC Check Solution No. 1 (PN 7120779, 25 mL; or 7120782, 125 mL)
• Deionized water

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 Perform the Tests | 93

• 5-mL Class A volumetric pipette

• 100-mL volumetric flask

Prepare Stock Solution:

1. Pipette a 5-mL aliquot of BioTek QC Check Solution No. 1 into a
100-mL volumetric flask.
2. Add 95 mL of DI water; cap and shake well.

Solution B

Required Materials:
• Deionized (DI) water
• FD&C Yellow No. 5 dye powder (typically 90% pure)
• Tween 20 (polyoxyethylene (20) sorbitan monolaurate) or BioTek wetting
agent, PN 7773002 (a 10% Tween solution)
• Precision balance with a capacity of 100 g minimum and readability of
0.001 g
• Weigh boat
• 1-liter volumetric flask

Prepare Stock Solution:

1. Weigh out 0.092 gram of FD&C No. 5 yellow dye powder into a weigh boat.
2. Rinse the contents into a 1-liter volumetric flask.
3. Add 0.5 mL of Tween 20, or 5 mL of BioTek’s wetting agent.
4. Make up to 1 liter with DI water, cap, and shake well.

96-well, flat-bottom microplates are required for the liquid tests

(Corning Costar #3590 is recommended). Use new microplates;
any fingerprints or scratches may cause variations in readings.

Before running the tests, ensure that the reader is not running in
Rapid Mode. To check this, select UTIL > READ and then cycle
through the prompts until READ IN RAPID MODE? is
displayed. Choose NO for an accurate result.

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Perform Liquid Test 1

1. Using freshly prepared stock solution A or B (the concentrated solution),

prepare a 1:2 dilution using DI water (one part stock, one part DI water; the
resulting solution is a 1:2 dilution).
2. Pipette 200 µL of the concentrated solution into Column 1 of the plate.
3. Pipette 200 µL of the diluted solution into Column 2 of the plate.

™ After pipetting the diluted test solution into the microplate and
before reading the plate, we strongly recommend shaking the plate
for four minutes. This will allow any air bubbles in the solution to
settle and the meniscus to stabilize. If a plate shaker is not
available, wait 20 minutes after pipetting the diluted test solution
before reading the plate.

4. Read the microplate five times at 405 nm using the Normal Read Mode,
single wavelength, no blanking (“Normal” plate position).
5. Without delay, rotate the microplate 180° so that well A1 is now in the H12
position. Read the plate five more times (“Turnaround” plate position).
6. Print the ten sets of raw data, or export the data to an Excel spreadsheet
using Gen5. The mathematical computations described below may then be
performed and the template kept for future tests.

Calculations:

7. Calculate the mean OD value for each well location in columns 1 and 2 for
the five plates read in the Normal position, and then again for the five
plates read in the Turnaround position. This will result in 32 mean values.
8. Perform a mathematical comparison of the mean values for each microwell
in its Normal and Turnaround positions (that is, compare A1 to H12, B1 to
G12, H1 to A12, and so on). To pass this test, the differences in the
compared mean values must be within the accuracy specification for the
instrument. For example:
If the mean value for well A1 in the Normal position is 1.902, where the
specified accuracy is ± 1.0% ± 0.010 OD, then the expected range for the
mean of the same well in its Turnaround (H12) position is 1.873 to 1.931
OD. 1.902 * 0.010 + 0.010 = 0.029; 1.902 – 0.029 = 1.873; 1.902 + 0.029 =
1.931

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™ If any set of mean values is out of the expected range, review the
other three sets of mean values for the same channel pair. For
example, if the A1/H12 comparison fails (the wells are not within
the expected range of each other), review the comparisons of
A2/H11, H1/A12, and H2/A11. If two or more sets of mean values
for a channel pair are out of the expected range, there is a
problem with one of the eight read channels. If only one of the
four mean values results in a failure, check the well for debris and
the plate for scratches or fingerprints.

Accuracy Specification:
For comparison in this test, the following accuracy specification is applied,
using Normal reading mode and a 96-well microplate.
± 1.0% ± 0.010 OD from 0.000 OD to 2.500 OD at 405 nm

Perform Liquid Test 2

™ The recommended method of testing the instrument performance is to

use the Absorbance Test Plate to confirm alignment, repeatability, and
accuracy, which will also confirm linearity. If a Test Plate is not
available, Liquid Test 2 can be used for these tests.

Required Materials:
• A new 96-well, flat-bottom microplate (Corning Costar #3590 is
recommended)
• Ten test tubes, numbered consecutively, stored in a rack
• Calibrated hand pipette (Class A volumetric pipette recommended)
• Stock solution A or B (these are the same solutions as for Liquid Test 1)
• A 0.05% solution of deionized water and Tween 20

Prepare Dilutions:
Create a percentage of dilution series, beginning with 100% of the original
concentrated stock solution (A or B) in the first tube, 90% of the original
solution in the second tube, 80% in the third tube, all the way to 10% in the
tenth tube. Dilute using the 0.05% solution of deionized water and Tween®
20. This solution can also be made by diluting the BioTek wetting agent 200:1.

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Test Tube Dilutions

Tube Number 1 2 3 4 5 6 7 8 9 10
Volume of Concentrated 20 18 16 14 12 10 8 6 4 2
Solution (mL)
Volume of 0.05% Tween 0 2 4 6 8 10 12 14 16 18
Solution (mL)
Absorbance expected 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2
if concentrated solution
is 2.0 at 200 µL

™ The choice of dilutions and the absorbance of the original solution can
be varied. Use this table as a model for calculating the expected
absorbances of a series of dilutions, given a different absorbance of
the original solution.

Prepare the plate:

1. Pipette 200 µL of the concentrated solution from tube 1 into each well of the
first column, A1 to H1, of the microplate.
2. Pipette 200 µL from each of the remaining tubes into the wells of the
corresponding column of the microplate (tube 2 into wells A2 to H2, etc.).

™ The choice of dilutions and the absorbance of the original solution

can be varied. Use the table on the previous page as a model for
calculating the expected absorbances of a series of dilutions, given
a different absorbance of the original solution.

™ After pipetting the diluted test solution into the microplate and
before reading the plate, we strongly recommend shaking the plate
for four minutes. This will allow any air bubbles in the solution to
settle and the meniscus to stabilize. If a plate shaker is not
available, wait 20 minutes after pipetting the diluted test solution
before reading the plate.

Perform the Linearity Test (Test A)

1. Read the prepared microplate five times using Normal mode, dual
wavelength at 450 nm with 630 nm as the blank.
™ Do not discard the plate; you will use it again for Test C.

2. Print the ten sets of raw data or export them to an Excel spreadsheet using
Gen5. The mathematical computations described below may then be
performed and the template kept for future data reduction.

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Calculations:
1. Calculate the mean absorbance for each well, and then average the means
for each concentration.
2. Perform a regression analysis on the data to determine if there is adequate
linearity. Since it is somewhat difficult to achieve high pipetting accuracy
when conducting linear dilutions, an R Square value of at least 0.99 is
considered adequate.

Calculate Repeatability (Test B)

1. Calculate the mean and standard deviation for the five readings taken
above at each concentration. Only one data set needs to be analyzed. The
well that shows the most variation for any concentration is selected for data
reduction.
2. For each mean below 2.500 OD, calculate the allowed deviation using the
repeatability specification for a 96-well plate of ± 0.5% ± 0.005 OD from
0.000 OD to 2.500 OD at 405 nm.
3. The standard deviation for each set of readings should be less than the
allowed deviation. For example:
Absorbance readings of 1.950, 1.948, 1.955, 1.952, and 1.950 will result in a
mean of 1.951, and a standard deviation of 0.0026. The mean (1.951)
multiplied by 0.5% (1.951 * 0.005) = 0.0098, which, when added to the 0.005
(0.0098 + 0.005) = 0.0148 OD, which is the allowable deviation. Since the
standard deviation is less than this value, the reader meets the test criteria.
Repeatability Specification:
± 0.5% ± 0.005 OD from 0.000 OD to 2.500 OD at 405 nm

Channel-to-Channel Variation and Alignment (Test C)

1. Using the plate prepared for Test A on the previous page, conduct a
Turnaround test by reading the plate with the A1 well in the H12 position
five times. This test results in two comparisons of each channel to its
corresponding channel.
2. Calculate the means of the wells in column 1 in the Normal plate position
(data is from Test A) and in the turnaround position (from Step 1 above).
Compare the mean reading for well A1 to its mean reading when in the
H12 position.
3. Next, compare the mean reading for well A1 to its mean reading when in
the H12 position. Next, compare the mean values for the other wells to
their corresponding mean values with the well in the turnaround position.
(Compare B1 to G12, C1 to F12, D1 to E12, E1 to D12, F1 to C12, G1 to B12,
H1 to A12). The difference in the values for any two corresponding wells

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98 | Chapter 4: Instrument Qualification

should be within the accuracy specification for the instrument. For

example:
If the mean of well A1 in the normal position is 1.902, where the specified
accuracy is ± 1.0% ± 0.010 OD, then the expected range for the mean of the
same well in the H12 position is 1.873 to 1.931 OD. (1.902 * 1.0% = 0.019 +
0.010 = 0.029, which is added and subtracted from 1.902 for the range.)
If any set of well values is out of the expected range, review the other set
for the same channel pair. Thus, if A1 and H12 are not within range of each
other, review the compliance of H1 to A12, A2 to H11, and H2 to A11. This
will confirm that there is a problem in one of the eight read channels, or
indicate that the result of one set of wells was in error. If any two sets of
well values for a channel pair are out of the allowed accuracy range, there
may be contamination on one of the lenses.
If the four corner wells are within the repeatability range, the reader is also
in alignment.

Perform Liquid Test 3 (“UV” Models Only)

™ Liquid Test 3 is an optional test offered for sites that must have proof
of linearity at wavelengths lower than those attainable with the
Absorbance Test Plate. This test verifies operation of the ELx808 at
340 nm and is optional because the reader has good “front-end”
linearity throughout its wavelength range.

Required Materials:
• 340 nm filter installed in the reader
• New 96-well, flat-bottom microplates (Corning Costar #3590 is
recommended)
• Calibrated hand pipette(s)
• Beakers and graduated cylinder
• Precision balance with readability to 0.01 g
• Buffer solution as described below

Buffer Solution

• Deionized water
• Phosphate-buffered saline (PBS), pH 7.2-7.6, Sigma tablets #P4417 (or
equivalent)
• β-NADH Powder (β-Nicotinamide Adenine Dinucleotide, Reduced
Form) Sigma bulk catalog number N 8129, or preweighed 10-mg vials,
Sigma number N6785-10VL (or BioTek PN 98233). Store the powder
according to the guidelines on its packaging.

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 Perform the Tests | 99

1. Prepare a PBS solution using the Sigma tablets.

2. In a beaker, mix 50 mL of the PBS solution with 10 mg of the β-NADH
powder and mix thoroughly. This is the 100% Test Solution.

Perform the Test

1. Check the absorbance of a sample of the 100% test solution at 340 nm on
the microplate reader. This solution will have an optical density
(absorbance) of approximately 0.700 to 1.000. This value is not critical, but it
should be within this absorbance range. If low, adjust up by adding
β -NADH powder until the high-level solution is at least at the lower end of
this range. Do not adjust if slightly high.
2. Prepare the 75% Test Solution by mixing 15 mL of the 100% Test
Solution with 5 mL of the PBS solution.
3. Prepare the 50% Test Solution by mixing 10 mL of the 100% Test
Solution with 10 mL of the PBS solution.
4. Pipette the three solutions into the new 96-well microplate:
¾ 150 µL of the 100% Test Solution into all wells of columns 1 and 2
¾ 150 µL of the 75% Test Solution into all wells of columns 3 and 4
¾ 150 µL of the 50% Test Solution into all wells of columns 5 and 6

™ After pipetting the diluted test solution into the microplate and
before reading the plate, we strongly recommend shaking the plate
for four minutes. This will allow any air bubbles in the solution to
settle and the meniscus to stabilize. If a plate shaker is not
available, wait 20 minutes after pipetting the diluted test solution
before reading the plate.

5. Read the microplate five times using Normal mode, single wavelength at
340 nm, no blanking (or blank on air).
6. Print the five sets of data, or export the data to an Excel spreadsheet using
Gen5. The mathematical computations described below may then be
performed and the template kept for future data reduction.

Calculate Repeatability
1. For each well, calculate the mean and standard deviation of the five
readings. Only one data set needs to be analyzed for each concentration.
The well that shows the most variation for each concentration is selected
for data reduction.
2. For each mean calculated in Step 1, calculate the allowed deviation using
the repeatability specification for a 96-well plate of ± 1.0% ± 0.005 OD from
0.000 to 2.000 OD at 340 nm.

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100 | Chapter 4: Instrument Qualification

3. For each well, compare the standard deviation calculation in Step 1 with
the allowed deviation calculation in Step 2. The standard deviation should
be less than the allowed deviation. For example:
Five readings in well A1 of 0.802, 0.802, 0.799, 0.798, and 0.801 will result in
a mean of 0.8004, and a standard deviation of 0.0018.
To calculate the allowed deviation for well A1, multiply the mean by 1.0%
and add 0.005 (0.8004 * 0.010 + 0.005) to get 0.013. Since the standard
deviation for well A1 is less than 0.013, the well meets the test criteria.
Repeatability Specification:
± 1.0% ± 0.005 OD from 0.000 to 2.000 OD at 340 nm

Calculate Linearity
1. For each of the three Test Solutions, calculate the mean absorbance for the
wells containing that solution (mean of wells A1 to H2, A3 to H4, and A5 to
H6).
2. Perform a regression analysis on the data to determine if there is adequate
linearity. Since it is somewhat difficult to achieve high pipetting accuracy
when conducting linear dilutions, an R Square value of at least 0.99 is
considered adequate.

BioTek Instruments, Inc.

Chapter 5

Preventive Maintenance

This chapter provides step-by-step instructions for maintaining the

ELx808 in top condition, to ensure that the reader continues to
perform to specification.

Recommended Maintenance Schedule ................................ 102 

Warnings and Precautions ................................................ 102 
Clean Exposed Surfaces ................................................... 103 
Inspect and Clean the Wavelength Filters ........................... 104 
(Optional) Lubricate Robotic Components ........................... 105 
Robotic Motor Adjustment Procedure ............................. 107 
Replace the Lamp and Clean the Contacts .......................... 107 
Decontamination ............................................................ 111 
Purpose .................................................................... 111 
Tools and Supplies...................................................... 112 
Decontamination Procedure ......................................... 112 
102 | Chapter 5: Preventive Maintenance

Recommended Maintenance Schedule

A general Preventive Maintenance (PM) regimen for all ELx808 models includes
periodically cleaning all exposed surfaces, inspecting/cleaning the wavelength filters,
replacing the lamp, and decontamination. Robotic models should have certain components
lubricated. The table below recommends Preventive Maintenance tasks and the frequency
with which each task should be performed.

™ The risk factors associated with your tests may require that some or
all of the procedures be performed more or less frequently than shown
below.

Frequency

Task Every Before

Every six
three As needed storage or
months
months shipment

Clean Exposed Surfaces, p. 103 9

Inspect/Clean Wavelength
9
Filters, p. 104

(Optional) Lubricate Robotic

9
Components, p. 105

Replace Lamp and Clean after ~500

Contacts, p. 107 hours

Decontamination, p. 111 9

Warnings and Precautions

Please read the following before performing any maintenance procedures.

Warning! Internal Voltage. Turn off and unplug the

instrument for all maintenance and repair operations.

Warning! Wear prophylactic gloves when handling

contaminated instruments. Gloved hands should be
considered contaminated at all times; keep gloved hands
away from eyes, mouth, nose, and ears. Eating and drinking
while decontaminating instruments is not advised.

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 Clean Exposed Surfaces | 103

Warning! Mucous membranes are considered prime entry

routes for infectious agents. Wear eye protection and a
surgical mask when there is a possibility of aerosol
contamination. Intact skin is generally considered an
effective barrier against infectious organisms; however, small
abrasions and cuts may not always be visible. Wear
protective gloves when handling contaminated instruments.

Important! Do not immerse the instrument, spray it with

liquid, or use a “wet” cloth. Do not allow water or other
cleaning solution to run into the interior of the instrument. If
this happens, contact BioTek’s Technical Assistance Center.

Important! Do not soak the keypad – this will cause

damage. Moisten a clean cloth with deionized or distilled
water and wipe the keypad. Dry immediately with a clean,
dry cloth.

Clean Exposed Surfaces

Important! Turn off and unplug the ELx808 for the

cleaning procedure.

Exposed surfaces may be cleaned (not decontaminated) with a cloth moistened (not
soaked) with water or water and a mild detergent. You will need:
• Mild detergent
• Deionized or distilled water
• Clean cotton cloths
To clean the exposed surfaces:
1. Turn off and unplug the instrument.
2. Moisten a clean cotton cloth with water, or with water and the mild detergent.
Do not soak the cloth!
3. Wipe the plate carrier, the inside of the plate carrier door, and all exposed
surfaces of the instrument.

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104 | Chapter 5: Preventive Maintenance

4. If detergent was used, wipe all surfaces with a cloth moistened with water.
5. Use a clean, dry cloth to dry all wet surfaces.

Inspect and Clean the Wavelength Filters

Important! To properly store interference filters for the

ELx808 during extended periods of non-use, package the
filters in a light-tight envelope or container, away from
high humidity and direct sunlight. This will ensure the
longest life for the filters. When handling the filters, keep
the surfaces clean from fingerprints and debris by simply
wiping with a lens tissue or other lint-free cloth.

Laboratory air is used to cool the lamp, and the wavelength filters can become dusty as a
result. The filters should be inspected and cleaned at least every three months. You will
need:
• Tape
• Screwdriver
• Isopropyl, ethyl, or methyl alcohol
• Lens-cleaning tissue
To inspect and clean the wavelength filters:
1. If you have not already done so, turn off and unplug the reader.
2. Temporarily tape the reader’s plate access door shut.
3. Using your fingertips, locate the seven perimeter screws around the sides and
front where the reader’s top shroud meets the base. Remove the screws using a
screwdriver.
™ Tip: Bring the reader to the edge of the work surface to access the
screws without having to turn the reader upside down.

4. Carefully raise the front of the shroud and tip it toward the back of the
instrument.
5. The filter wheel is located in the left-rear quadrant inside the reader. Remove
the filter wheel by pulling it off its magnetic hub.
6. Inspect the glass filters for speckled surfaces or a halo effect. This may indicate
deterioration due to moisture exposure over a long period of time. If you have
any concerns about the quality of the filters, contact your BioTek
representative.

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 (Optional) Lubricate Robotic Components | 105

7. Clean the filters using lens-cleaning tissue moistened with a small amount of
isopropyl, ethyl, or methyl alcohol. Ensure that the filters remain in their
current locations.
8. Replace the filter wheel on its magnetic hub. Ensure that the peg on the wheel
lines up with the notch on the hub, and that the filter wheel is positioned flat
against the hub and rotates freely.
9. Close the top shroud and replace the seven perimeter screws.

™ Store filters in a dry environment, such as a desiccator, if they will not

be used for an extended period.

™ If the reader continues to fail tests in Chapter 4, Instrument

Qualification, after you have cleaned the filters and the lamp
contacts, or if one or more filters appear to have deteriorated, contact
your BioTek Representative or BioTek.

(Optional) Lubricate Robotic Components

This section applies to ELx808 robotic models only. You will need lubricant, BioTek
PN 66039.
1. Remove the top shroud of the instrument (see instructions in Chapter 2).
2. Lubricate the components as shown in Figure 8a and Figure 8b every six
months, or after 10,000 cycles, according to the instructions in the table below.

Component
Location Procedure

A Lubricate the underside of the bracket.

B Lightly lubricate the shaft indicated.

C Apply a heavy coating of lubricant to both sides of the motor

shaft. Run the motor shaft back and forth through the motor to
ensure that the internal drive nut is heavily lubricated. Use the
robotic motor adjustment procedure described on the next
page to work the lubricant into the internal drive nut.

D Lightly lubricate the surface of the roller.

E Lightly lubricate the underside of the hook-in bracket.

3. Reattach the top shroud.

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106 | Chapter 5: Preventive Maintenance

Figure 8a: Location of bracket, motor shaft, and main PCB/motor assembly

Figure 8b: Roller surface and hook-in bracket

BioTek Instruments, Inc.

 Replace the Lamp and Clean the Contacts | 107

Robotic Motor Adjustment Procedure

After lubrication/reassembly, the robotic door needs to be set to the open and closed
positions. Press the following keys to adjust the door.
At the Main Menu:
1. Press the UTIL soft key.
2. Press the SETUP soft key.
3. Press the hidden key between the Main Menu and Previous Screen keys.
You should see the Door Adjustment screen.
4. Press the CLOSE soft key.
5. Press the UP soft key until you just see the door move up. You may have to
press the key many times before the door moves. When the door does
move, press the DOWN soft key 4 times. This positions the door lift ram
just behind the door roller in the closed position.
6. Press the OPEN soft key.
7. Press the UP soft key to raise the door to the desired height. Press the
DOWN soft key if you want to adjust the door downward.

Important! Do not open the door to its

maximum open position.

8. Press the Main Menu key.

9. Press the CLOSE soft key. This will close the door. The door should rest
closed on the top cover.

Replace the Lamp and Clean the Contacts

™ There are two types of lamp assemblies. Steps 7 and 8 only pertain to
the newer assembly. Review the photos below to determine which
instructions apply to your reader.

The reader’s lamp, on average, should provide 500 hours of total service before it needs to
be replaced. This equates to approximately six months to one year of use under routine
conditions. The intensity of the lamp will slowly drop over time until the reader’s run-time
self-check detects a low signal and warns the user with a displayed error code. The bulb
should be replaced at this time (PN 3400508), and the electrical contacts in the lamp socket
cleaned.

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108 | Chapter 5: Preventive Maintenance

Warning! Hot surface. The lamp is hot when the instrument

is turned on. Turn off and unplug the reader. Allow the lamp
to cool down before attempting to replace it.

Important! Oxide coating on the electrical contacts can

reduce lamp output. Clean the contacts in the socket
whenever the lamp is replaced (approximately every 500
hours).

You will need:

• Tape
• Screwdriver
• Cotton swabs or pencil eraser
• Isopropyl alcohol
• Lamp, BioTek PN 3400508

To replace the lamp:

1. Turn off and unplug the reader. Allow the lamp to cool for at least ten minutes.
2. Temporarily tape the reader’s plate access door shut.
3. Using your fingertips, locate the seven securing (perimeter) screws around the
sides and front where the reader’s top shroud meets the base. Remove the
screws using a screwdriver.
™ Tip: Bring the reader to the edge of the work surface to access the
screws without having to turn the reader upside down.

4. Carefully raise the front of the shroud and tip it toward the back of the
instrument.
5. The lamp is located in the left rear quadrant inside the reader. Unplug the
lamp connector from the circuit board.

Important! Do not touch the glass lens in the lamp

bracket.

6. The lamp bracket is attached to the side of the instrument with two screws,
which are accessed from the outer left side of the instrument. Using a
screwdriver, loosen (do not remove) these two screws. Slide the bracket out of
the instrument.

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 Replace the Lamp and Clean the Contacts | 109

7. (Lamp Assembly 1 Only) Carefully unscrew the two thumbscrews on the

lamp clamp that holds the bulb in the bracket. Remove the screws, springs,
washers, and lamp clamp and set them aside.
8. (Lamp Assembly 1 Only) Slide the rubber O-ring down the bulb socket away
from the bulb.
9. Remove the bulb from its socket by pushing the socket and the bulb together
and rotating the socket counterclockwise.
10. Clean the two contacts in the socket, using a cotton swab moistened with
isopropyl alcohol, and allow them to dry for a few minutes. An alternative
cleaning technique would be to rub the contacts with a pencil eraser.

To install the lamp, reverse steps 1 through 9. Keep your fingers out of the reflector
interior and away from the bulb. Ensure that the bulb is set flat against its housing
before reinstallation in the instrument.

O-ring lamp clamp

washers

thumbscrews
springs

Figure 9: New lamp hardware for the ELx808 (Lamp Assembly 1)

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110 | Chapter 5: Preventive Maintenance

Figure 10: New lamp hardware for the ELx808, assembled (Lamp Assembly 1)

Figure 11: Location of the lamp in the ELx808 (Lamp Assembly 2)

BioTek Instruments, Inc.

 Decontamination | 111

Decontamination

Purpose

Any laboratory instrument that has been used for research or clinical analysis is
considered a biohazard and requires decontamination prior to handling.
Decontamination minimizes the risk to all who come in contact with the instrument
during shipping, handling, and servicing. Decontamination is required by the U.S.
Department of Transportation regulations.
Persons performing the decontamination process must be familiar with the basic setup
and operation of the instrument.

Important! BioTek Instruments, Inc. recommends the use of

the following decontamination solutions and methods based on
our knowledge of the instrument and recommendations of the
Centers for Disease Control and Prevention (CDC). Neither
BioTek nor the CDC assumes any liability for the adequacy of
these solutions and methods. Each laboratory must ensure that
decontamination procedures are adequate for the Biohazard(s)
they handle.

Warning! Internal Voltage. Turn off and unplug the

instrument for the decontamination procedure.

Warning! Wear prophylactic gloves when handling

contaminated instruments. Gloved hands should be
considered contaminated at all times; keep gloved hands away
from eyes, mouth, nose, and ears. Eating or drinking while
decontaminating instruments is not advised.

Warning! Mucous membranes are considered prime entry

routes for infectious agents. Wear eye protection and a surgical
mask when there is a possibility of aerosol contamination.
Intact skin is generally considered an effective barrier against
infectious organisms; however, small abrasions and cuts may
not always be visible. Wear protective gloves when performing
the Decontamination procedure.

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112 | Chapter 5: Preventive Maintenance

Tools and Supplies

• Sodium hypochlorite (NaClO, or bleach)

• 70% isopropyl alcohol (alternative to bleach)
• Deionized or distilled water
• Safety glasses
• Surgical mask
• Protective gloves
• Lab coat
• Biohazard trash bags
• 125-mL beakers
• Clean cotton cloths

Decontamination Procedure

Warning! The bleach solution is caustic; wear gloves and

eye protection when handling the solution.

Do not immerse the instrument, spray it with liquid, or use

a “wet” cloth. Do not allow the cleaning solution to run into
the interior of the instrument. If this happens, contact the
BioTek Service Department.

Do not soak the keypad – this will cause damage.

Important! Turn off and unplug the instrument for all

decontamination and cleaning operations.

1. Turn off and unplug the instrument.

2. Prepare an aqueous solution of 0.5% sodium hypochlorite (bleach). As an
alternative, 70% isopropyl alcohol may be used if the effects of bleach are a
concern.
• Be sure to check the percent NaClO of the bleach you are using; this
information is printed on the side of the bottle. Commercial bleach is
typically 10% NaClO; if this is the case, prepare a 1:20 dilution. Household
bleach is typically 5% NaClO; if this is the case, prepare a 1:10 dilution.

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 Decontamination | 113

3. Moisten a clean, lint-free cloth with the bleach solution or alcohol. Do not
soak the cloth! Wipe down the carrier and all exposed instrument surfaces
with the bleach solution.
• Wipe the keypad (do not soak). Wipe again with a clean cloth moistened
with deionized or distilled water. Dry immediately with a clean, dry cloth.
• Wipe the plate carrier, the inside of the carrier door, and all exposed
surfaces of the instrument.
4. Wait 20 minutes. Moisten a cloth with deionized or distilled water and wipe all
surfaces of the instrument that have been cleaned with the bleach solution or
alcohol.
5. Use a clean, dry lint-free cloth to dry all wet surfaces.
6. Reassemble the instrument as necessary.
7. Discard the used gloves and cloths, using a Biohazard trash bag and an
approved Biohazard container.

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114 | Chapter 5: Preventive Maintenance

BioTek Instruments, Inc.

Chapter 6

Troubleshooting and Error

Codes

This chapter provides a list of Error Codes that may appear on the
ELx808 or in Gen5, and their probable causes.

Overview ....................................................................... 116

Diagnostics................................................................ 116
Terms Used in the Error Codes Tables ........................... 116
Error Codes (General) ..................................................... 117
Error Codes (Fatal) ......................................................... 121
116 | Chapter 6: Error Codes

Overview
An error code that appears on the display of the ELx808 (during instrument control of the
reader), or in Gen5 software (during PC control of the reader) is displayed as a four-digit
identifier. The first digit will be 0, 1, 2, or A.
• 0, 1, or 2 denote a noncritical or “General” error, which means that it is still
possible for the ELx808 to communicate with the controlling software and run
the reader system test. See Error Codes (General) beginning on page 117.
• A denotes a more serious error with the memory or processing, which requires
that the ELx808 be turned off and then powered up before any diagnostics can
be performed. If the error reappears, call BioTek TAC for troubleshooting
assistance (see Chapter 1). See Error Codes (Fatal) on page 121.

Diagnostics

If an error code is displayed, a System Self-Test should be conducted for diagnostic

purposes. Refer to the following brief instructions for performing the Self-Test:
• From the Main Menu screen on the ELx808 display, select UTIL > TESTS
> SYSTEM.

• In Gen5, access the System menu and select Diagnostics > Run System
Test.

Terms Used in the Error Codes Tables

• Air Blank: A full light reading through a filter with no plate in the light path.

• Dark Current: A reading taken with the light blocked to measure background
light levels in the reading chamber. Also used as a measure of background
electronic noise within the measurement circuit.
• Gain: An automatic electronic adjustment to the measurement circuit. The
gain adjustment compensates for changing light levels or filter variations. For
example, if the lamp output decreases slightly, the gain will increase to make
up the difference.
• Axis: Refers to a motor that positions the filter wheel or plate carrier.

• Offset: A numerical limit, usually a range. For example, if the gain fails an
offset test, it may be too high or too low.

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 Error Codes (General) | 117

Error Codes (General)

Code Probable Cause

0200 Plate carrier did not find the home sensor.
0201 Filter wheel motor did not find the home sensor; filter wheel not
installed or detached from hub.
Errors 0200 and 0201 indicate that a motor was not able to move to its
“home” position as registered by feedback from an optical sensor.
Probable causes:
• Carrier movement may be limited so that the optical sensor cannot be
interrupted. Check the carrier path for obstructions.
• Filter wheel not installed. The wheel is shipped in a separate envelope
and may not be installed or may have broken free of the magnetic
hub.
• Filter wheel movement may be impeded. Remove the wheel to confirm
if a loose filter is impeding the movement of the wheel.
0300 Carrier failed to find light beam.
0301 Filter wheel did not find home.
Errors 0300 and 0301 indicate that a particular axis has moved to a point
where the light beam from the optics is no longer detectable by the
measurement electronics. This error is usually only seen during the service
Auto Cal sequence.
Probable causes:
• Carrier: Lamp may be out or blocked by a faulty filter wheel. A loose
belt or a loose motor pulley may cause the carrier to ignore movement
instructions.
• Filter Wheel Motor: The filter wheel is missing or loose or a loose filter
may be impeding filter wheel movement.
0400 Carrier x-axis failed position verify.
0401 Filter Wheel motor failed position verify.
Errors 0400 and 0401 indicate that an axis failed its Position Verify test. After
moving a predefined number of steps from a home position, the motor should
return to the home position in the proper amount of time and steps. If the
axis moves back to its home position in the wrong amount of time or in too
few or too many steps, the test fails.
Probable causes:
• Mechanical obstruction.
• Carrier belt or pulley slipping.
• Loose filter wheel hub.
• Loose filter may be impeding filter wheel movement.
• Defective motor drive circuit or 12-volt power problem.

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118 | Chapter 6: Error Codes

Code Probable Cause

0500 Filter is not installed.
0502 to 0500 indicates an empty filter location on the filter wheel, or that the filter
0506 wheel is not installed.
Probable causes:
• All filter locations must have either a filter or a filter blank (BTI
PN 3122037) installed. The reader may also display this error if the
filter wheel is not installed.
0601 to Filters 1 through 6 Gain out of range.
0606 Errors 0601 through 0606 indicate that the gain for a specific filter is out of
range. Gain out of range indicates that the lamp is out, or that a particular
filter absorbs or transmits entirely incorrect light levels.
Probable causes:
• Lamp out. Check that the lamp is lit when the unit is ON. Replace lamp
if necessary.
• A missing, defective, or misplaced filter or filter blank will cause this
error. Observe that the filters in the wheel match the filters
programmed in the software. Look for obvious physical defects in the
filters.
0700 Reader failed noise test.
Error 0700 indicates significant variations in background electronic noise were
detected when blocking the light and increasing the Gain to maximum.
Probable causes:
• Electrical noise penetrating the measurement chamber. The bottom
and top shrouds are part of the electrical shielding. Check that the
shrouds are installed and properly fastened.
• Ambient light leak. Check that the plate chamber door is closed to
keep out ambient light.
• Missing or loose filter allowing unintended lamp light into the
measurement chamber. Check filter wheel for empty positions.
• Internal electronic noise caused by a faulty Analog PCB or faulty
internal grounding.
0800 Reader failed offset test.
Error 0800 indicates electronic signal detected is outside of acceptable limits
at maximum gain when blocking the light.
Probable causes:
• Electrical noise penetrating the measurement chamber. The bottom
and top shrouds are part of the electrical shielding. Check that the
shrouds are installed and properly fastened.
• Ambient light leak. Check that the plate chamber door is closed to
keep out ambient light.
• Missing or loose filter allowing unintended lamp light into the
measurement chamber. Check filter wheel for empty positions.
• Internal electronic noise caused by a faulty Analog PCB or faulty
internal grounding.

BioTek Instruments, Inc.

 Error Codes (General) | 119

Code Probable Cause

0900 Read time dark value out of range.
Error 0900 indicates that the dark current value taken during the current read
is significantly different than the same reading during the power-up self-
check.
Probable cause:
• Read chamber door has opened or closed relative to when the unit was
powered ON. Close door and reboot reader to correct.
• Electrical noise penetrating the measurement chamber. The bottom
and top shrouds are part of the electrical shielding. Check that the
shrouds are installed and properly fastened.
• Internal electronic noise caused by a faulty Analog PCB or faulty
internal grounding.
0A00, Read time air blank out of range.
0A01 Error 0A00 indicates that the blank (full signal) reading taken during the
to 0A05 current read has changed significantly from the same reading taken during
the power-up self-check.
Probable cause:
• Lamp has failed since power-up. Check that the lamp is lit and replace
if necessary.
• Analog PCB failure.
0C00 Printer time out error.
Error 0C00 indicates that the time allotted for the instrument to make a valid
connection to a printer has expired.
Probable cause:
• Printer out of paper, not connected, or not on-line.
0D00 Reader failed calibration checksum test.
Internal software corruption.
0E00, Wavelength not detected in reader’s filter table.
0E01 to Compare filters’ wavelengths to the filter table.
0E05
0F00 to Reader’s filter or channel signal is out of specified range.
OF86 Check for lamp failure, and/or replace lamp.
1000 Required reader configuration data missing.
Re-download software basecode and assays.
1100 Failed configuration checksum test.
Re-download software basecode and assays.
1200 Calibration data missing.
1201 Calibration value missing
Filter table was updated but System Test was not run. Run a system test.
1300 Motor not homed.
Autocal error, see error 0200.

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120 | Chapter 6: Error Codes

Code Probable Cause

1400 Assay incubation error.
Assay requires incubation but the instrument does not appear to have an
incubator.
1500 Incubator failed to hold temperature within tolerances during the
assay.
1600 Computer control assay definition error.
This error is used only for software development.
1700 Kinetic interval too short for selected options.
1800 Too many kinetic intervals selected.
2311 to Lamp unstable.
2386 Problem with lamp output during power-up or while reading a plate. If a 23xx
error is displayed during a System Test or during startup, the last digit
indicates the filter wheel position. If a 23xx error is displayed during a read,
the last digit indicates the filter number in the list of filters used for the read.
Probable causes:
• Defective or old lamp; replace the lamp.
• Lamp not sitting correctly in lamp holder (always remove the holder to
install the lamp, then insert the holder with the lamp back into the
reader).
• Lamp contacts dirty. Clean with ethanol (refer to Chapter 5,
Preventive Maintenance).
• Lamp cable needs upgrade.
• Filter degradation.
• Filter not seated properly in filter wheel.
• Lower lenses for all or some channels failing and need replacement.
• Electrical noise or grounding issue.
• Shortened lamp life from performing long kinetic runs every day or
leaving the reader on 24 hours a day.
• When using 340 nm filters, the analog gain must be increased.

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 Error Codes (Fatal) | 121

Error Codes (Fatal)

These errors indicate conditions that require immediate attention. If a fatal error is
displayed, contact BioTek’s Technical Assistance Center for further instructions (refer to
the Product Support & Service section in Chapter 1).

Code Description
A100 Task control block not available.
A200 Read already in progress.
A300 <Device> not available.
A400 Failed code checksum test on powerup.
A500 DR steps alloc/free error < assay num>.
A600 Data flash write timed out.
A700 Data flash readback did not match write (test) <chip>.
A800 Code flash write timed out.
A900 Memory Manager corruption detected. Power instrument off, then back
on.

ELx808 Operators’ Manual

122 | Chapter 6: Error Codes

BioTek Instruments, Inc.

Appendix A

Sample Reports

The following pages contain examples of reports that can be

generated and/or printed from the ELx808 when it is running in
standalone mode. For details on how to print these reports, refer to
OUTPUT Options in Chapter 2, Installation. In addition, an Assay
List, Assay Definition, Map, and Result can be printed by choosing
REPORT from the Main Menu screen.

L
Important! The 'OUT' indication on reports means the OD for
an individual well, or the average OD for a group of wells, falls
outside the minimum/maximum OD range defined for the
assay. For the Quick Read assay on the ELx808, this range is -3.0
OD to +3.0 OD. For assays defined onboard by a user (Assay02-
Assay55), it is -4.0 OD to +4.0 OD.

Samples with Calls on Matrix Report .................................. 124 

Curve Fit Report ............................................................. 125 
Samples with Calls on Column Report ................................ 126 
Column Report without Samples ....................................... 127 
Panel Report .................................................................. 128 
Assay Detail Report ......................................................... 129 
Assay List ...................................................................... 130 
124 | Appendix A: Sample Reports

Samples with Calls on Matrix Report

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 Curve Fit Report | 125

Curve Fit Report

ELx808 Operator’s Manual

126 | Appendix A: Sample Reports

Samples with Calls on Column Report

BioTek Instruments, Inc.

 Column Report without Samples | 127

Column Report without Samples

ELx808 Operator’s Manual

128 | Appendix A: Sample Reports

Panel Report

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 Assay Detail Report | 129

Assay Detail Report

ELx808 Operator’s Manual

130 | Appendix A: Sample Reports

Assay List

Assay List
Version: 2.82
1 _Quick Read
2 ASSAY 02
3 ASSAY 03
4 ASSAY 04
5 ASSAY 05
6 ASSAY 06
7 ASSAY 07
8 ASSAY 08
9 ASSAY 09
10 ASSAY 10
11 ASSAY 11
12 ASSAY 12
13 ASSAY 13
14 ASSAY 14
15 ASSAY 15
16 ASSAY 16
17 ASSAY 17
18 ASSAY 18
19 ASSAY 19
20 ASSAY 20
21 ASSAY 21
22 ASSAY 22
23 ASSAY 23
24 ASSAY 24
25 ASSAY 25
26 ASSAY 26
27 ASSAY 27
28 ASSAY 28
29 ASSAY 29
30 ASSAY 30

BioTek Instruments, Inc.

Appendix B

Instructions for Programming a

New Assay

This appendix provides two sample assay kit instructions with step-by-
step instructions for programming each assay: one with a ratio
transformation calculation and a POS/NEG cutoff determination, and
another with a standard curve.

These examples are based on real assays; however, the results have not
been verified on any clinical kits. The kit instructions are provided so that
users can see how it is possible to translate the kit wording into an
ELx808 assay program. For clarity, only the user menu choices from the
reader screens are shown. See Chapter 3 for details.

Important! The user is responsible for programming the

reader properly according to their specific kit instructions, and
for verifying that the calculations are performed correctly.

Sample ANA Screen Enzyme Immunoassay Kit ........................ 132

Sample Anticardiolipin IgG Enzyme Immunoassay Kit ............... 136
132 | Appendix B: Programming a New Assay

Sample ANA Screen Enzyme Immunoassay Kit

This sample demonstrates the use of transformations and cutoffs.

Intended Use

This assay is designed for the in vitro screening of serum for the presence of specific
IgG antinuclear antibodies (ANAs), to aid in diagnosing certain systemic rheumatic
diseases. Sufficient materials are supplied to allow a maximum of 93 samples to be
screened in single, with a positive, cutoff, and negative controls.

Background

Antinuclear antibodies occur in a large number of patients with systemic rheumatic

diseases. These diseases are characterized by the presence of one or more ANAs.
Sera positive on this ANA kit should be tested for the specific autoantibodies
indicative of the various systemic rheumatic diseases.

Principle of the Assay

Microwells are pre-coated with purified antigens. The prediluted controls and diluted
patient samples are added to the wells, and autoantibodies recognizing one or a
combination of antigens bind during the first incubation. After washing the wells to
remove all unbound proteins, IgG conjugate is added. The conjugate binds to the
captured autoantibody, and the excess unbound conjugate is removed by a further
wash step. Substrate is added that causes a blue reaction, thereby exposing the bound
conjugate, and producing an intensity proportional to the concentration of
autoantibody in the sample. Phosphoric acid is added to each well to stop the reaction.
This produces a yellow end-point color, which is read at 450 nm.

Materials Supplied

• Instruction Leaflet: Giving full assay details.

• QC Certificate: Indicating the expected performance of the batch.
• ANA Coated Wells: 12 x 8 well strips coated with purified antigens.
• Type III Wash Buffer 20x Concentrate: 1 bottle containing 50 mL of a concentrated
buffer for washing the wells.
• Type III Sample Diluent: 2 bottles containing 50 mL of buffer for sample dilution.
Ready to use.
• ANA Positive Control: 1 bottle containing 1.8 mL of diluted stabilized serum.
Ready to use.

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 Sample ANA Screen Enzyme Immunoassay Kit | 133

• ANA Cutoff Control: 1 bottle containing 1.8 mL of diluted stabilized serum. Ready
to use.
• ANA Negative Control: 1 bottle containing 1.8 mL of diluted stabilized serum.
Ready to use.
• ANA Conjugate: 1 bottle containing 12 mL of peroxidase labeled antibody to IgG.
Colored red, ready to use.
• TMB Substrate: 1 bottle containing 14 mL TMB substrate. Ready to use.
• Stop Solution: 1 bottle containing 14 mL of 3M Phosphoric acid. Ready to use.

Additional Materials and Equipment – Not Supplied

• Automatic Microplate Plate Washer: This is recommended; however, plate

washing can be performed manually.
• Microplate Reader: Capable of measuring optical densities at 450 nm referenced on
air.
• Distilled or Deionized Water: This should be of the highest quality available.
• Calibrated Micropipettes: For dispensing 1000, 100, and 10 µL.
• Multichannel Pipette: Recommended for dispensing 100 µL volumes of conjugate,
substrate, and stop solution.
• Glass/Plastic Tubes: For sample dilution.

Quality Control

For an assay to be valid, all the following criteria must be met:

• Cutoff as well as Positive and Negative controls must be included in each run.
• The OD of the cutoff and the ANA result of the Negative and Positive Controls
should be in the ranges specified on the QC Certificate.
• For example: The absorbance of the Positive Control must be greater than
1.200 OD. The absorbance of the Negative Control must be less than 0.300 OD.

™ If the above criteria are not met, the assay is invalid and the test
should be repeated.

Calculation of the Sample Results

Use the following formula to calculate the ANA result for each sample:
Control or sample OD
x 10 = Control or sample value (U/mL)
Cutoff control OD

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134 | Appendix B: Programming a New Assay

Expected Values

The normal range was determined on serum from 200 normal adult blood donors. The
cutoff control has been set at a point equivalent to the upper normal limit, or a cutoff
level of 10 U/mL.
The ranges are provided as a guide only. ELISA assays are very sensitive and capable
of detecting small differences in sample populations. It is recommended that each
laboratory determine its own normal range, based on the population techniques and
equipment employed.

ANA Result Interpretation

< 10.0 Negative

> 10.0 Positive

Programming the Assay on the ELx808

From the Main Menu, press DEFINE. Enter the assay number and edit the name if
desired. At the DEFINE menu:
STEP COMMENTS
To program the reading method, press:
METHOD:
READ TYPE (Endpoint, Kinetic, or Scan):
Endpoint
DELAY FIRST READ:
If incubator model: INCUBATION TEMP
(Ambient or Temperature):
WAVELENGTH (Single or Dual): Single
MEASURE (Wavelength[s] to use): 450
SHAKE (First, Every, or None):
To program the plate map, press:
MAP
AUTO “Auto” mapping is normally preferred,
because it fills in the well IDs logically
and automatically after determining
which direction to map and how many
wells to fill.
DOWN Maps the wells down the column.
DOWN Locates replicates in a vertical
orientation down the column.
A01 Begins mapping at well location A01.
BLANK MAP: AIR Choose to blank on “AIR” if no blank
wells are required.
NUMBER STDS: 00

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 Sample ANA Screen Enzyme Immunoassay Kit | 135

STEP COMMENTS
REUSE STANDARD CURVE? Can only be programmed in assays
31-55. Cannot be reused on panels.
NUMBER CTLS: 03
CONTROL 1: PC
CONTROL 2: CTL1 Suggested for the cutoff control.
CONTROL 3: NC
NUMBER OF REPLICATES
PC: 01
CTL1: 03
NC: 01
SAMPLES: 91 User-defined; recommendation is to
fill the entire plate.
SAMPLE REPLICATES: 01
Use the MAP and MATH keys to
create the control validation formula:
FORMULA
VAL
CONTROL: When defining control validation
PC;x > 1.200 formulas, use “PC” to indicate the
criterion for each of the PC replicates,
NO. OF REPLICATES: 01 and “PC;x” to indicate the average of
NC;x < 0.300 the Positive Control replicates.
NO. OF REPLICATES: 01
To create the plate transformation for CALC, divide the ANA result by a cutoff standard.
FORMULA
*MORE
TRANS - VAR
SCOPE VARIABLE (SMP or OD) Select OD to advance to the formula
definition screen and define the
transformation variable (Tvar)
FORMULA
CTL1;X Defining TRANS VAR = CTL1;X
isolates the OD value for CTL1;X for
use in transformation.
TRANS:
FORMULA: (OD/TVAR)*10 Converts all OD values on plate to
”ANA Result” per the kit instructions.
To define a cutoff formula for Positive and Negative calls:
FORMULA
CUTOFF: 10.0
GREYZONE: 00%
SAMPLE > CUTOFF: POS

ELx808 Operator’s Manual

136 | Appendix B: Programming a New Assay

Sample Anticardiolipin IgG Enzyme Immunoassay

Kit

This sample demonstrates the use of standard curve and cutoffs.

Intended Use

This assay is intended for the in vitro measurement of IgG anticardiolipin antibodies in
serum, as an aid in the diagnosis of antiphospholipid syndrome (APS). Sufficient
materials are supplied to allow a maximum of 41 samples to be tested in duplicate or
89 in single, with a standard curve and positive and negative controls.

Background

Anticardiolipin antibodies are found in a wide range of conditions either transiently, in

some infectious diseases, or more persistently in autoimmune diseases such as
systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS).
Anticardiolipin antibodies have also been associated with a range of clinical conditions
including fetal loss, endocarditis, stroke, heart attack and autoimmune haemolytic.

Principle of the Assay

Microwells are pre-coated with cardiolipin and cofactor. Standards, controls, and
patient samples are added to the wells, and autoantibodies recognizing cardiolipin
bind during the first incubation. After washing the wells to remove all unbound
proteins, conjugate is added. The conjugate binds to the captured antibody, and the
excess unbound conjugate is removed by a further wash step. Substrate is added that
causes a blue reaction, thereby exposing the bound conjugate and producing an
intensity proportional to the concentration of autoantibody in the sample. Phosphoric
acid is added to each well to stop the reaction. This produces a yellow end-point color,
which is read at 450 nm.

Materials Supplied

• Instruction Leaflet: Giving full assay details.

• QC Certificate: Indicating the expected performance of the batch.
• Cardiolipin Coated Wells: 12 break-apart 8-well strips coated with bovine
cardiolipin antigen. The plate is packaged in a re-sealable foil bag containing two
desiccant pouches.
• Type II Sample Diluent: 2 bottles containing 50 mL of buffer for sample dilution.
Colored yellow, ready to use.

BioTek Instruments, Inc.

 Sample Anticardiolipin IgG Enzyme Immunoassay Kit | 137

• Type II Wash Buffer (20x Concentrate): 1 bottle containing 50 mL of a 20-fold

concentrated buffer for washing the wells.
• Cardiolipin IgG Standards: 5 bottles each containing 1.2 mL of diluted serum, with
the following concentrations of anticardiolipin autoantibody: 100, 50, 25, 12.5, 6.25
GPL U/mL. Ready to use.
• The standard set is calibrated against the Louisville APL reference preparation (see
Assay Calibration, page 138).
• Cardiolipin IgG Positive Control: 1 bottle containing 1.2 mL of diluted serum. The
expected value is given on the QC certificate. Ready to use.
• Cardiolipin Negative Control: 1 bottle containing 1.2 mL of diluted serum. The
expected value is given on the QC certificate. Ready to use.
• Cardiolipin IgG Conjugate: 1 bottle containing 12 mL of purified peroxidase
labeled antibody. Colored red, ready to use.
• TMB Substrate: 1 bottle containing 14 mL TMB substrate. Ready to use.
• Stop Solution: 1 bottle containing 14 mL of 3M phosphoric acid. Ready to use.

Additional Materials and Equipment – Not Supplied

• Automatic Microplate Plate Washer: This is recommended; however, plate

washing can be performed manually.
• Microplate Reader: Capable of measuring optical densities at 450 nm referenced on
air.
• Distilled or Deionized Water: This should be of the highest quality available.
• Calibrated Micropipettes: For dispensing 1000, 100, and 10 µL.
• Multichannel Pipette: Recommended for dispensing 100 µL volumes of conjugate,
substrate, and stop solution.
• Glass/Plastic Tubes: For sample dilution.

Quality Control

For an assay to be valid, all the following criteria must be met:

• Standards and the positive and negative controls must be included in each run.
• The values obtained for all the controls should be in the ranges specified on the
QC Certificate.
• The curve shape should be similar to the standard curve, shown on the QC
Certificate.
• If the above criteria are not met, the assay is invalid and the test should be
repeated.

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138 | Appendix B: Programming a New Assay

Calculate Mean Optical Densities (for assays run in duplicate only)

For each standard, control, and sample, calculate the mean OD of the duplicate
readings. The user must verify that the percentage coefficient of variation (%CV) for
each duplicate OD is less than 15.0%.

Plot Calibration Curve

The calibration curve can be plotted either automatically or manually as follows by

plotting the anticardiolipin autoantibody concentration on the log scale against the OD
on the linear scale for each calibrator:
• Automatic - Use appropriately validated software, and the curve fit that best
fits the data.
• Manual - Using log/linear graph paper, draw a smooth curve through the
points (not a straight line or point to point).

Treatment of Anomalous Points

If any one point does not lie on the curve, it can be removed. If the absence of this
point means that the curve has a shape dissimilar to that of the sample calibration
curve, or more than one point appears to be anomalous, then the assay should be
repeated.

Calculation of Autoantibody Levels in Controls and Samples

Read the level of the anticardiolipin autoantibody in the controls and diluted samples
directly from the calibration curve. The control values should fall within the range
given on the QC Certificate.

™ The standard values have been adjusted by a factor of 100 to

account for a 1:100 sample dilution. No further correction is
required.

Assay Calibration

The assays are calibrated against the Louisville reference LAPL-GM-100. One GPL unit
is defined as the cardiolipin binding activity of 1 µg/mL of an affinity purified IgG
anticardiolipin preparation from a standard serum.
The Louisville reference center recommends the following positive discrimination
criteria according to the recommendation of the 2nd International Anticardiolipin
Workshop.

BioTek Instruments, Inc.

 Sample Anticardiolipin IgG Enzyme Immunoassay Kit | 139

Criteria Range (GPLU/mL)

High Positive > 80

Medium Positive ≥ 20-80

Low Positive ≥ 10, < 20

Results Interpretation

The association between low positive levels of anticardiolipin antibodies and clinical
findings is unclear.
Normal population studies indicate that there is a higher prevalence of IgM positives
in the normal population than IgG, 9.4% and 6.5%, respectively. In normal pregnancy,
the levels are higher still at 17.0% (IgM) and 10.6% (IgG).

Expected Values

The normal range was determined on serum from 102 normal adult blood donors. The
ranges below are provided as a guide only. ELISA assays are very sensitive and
capable of detecting small differences in sample populations. It is recommended that
each laboratory determine its own normal range, based on the population techniques
and equipment employed.

lgG Anticardiolipin

< 11 GPL U/mL Negative result

> 11 GPL U/mL Positive result

Programming the Assay on the ELx808

From the Main Menu, press DEFINE. Enter the assay number and edit the name if
desired. At the DEFINE menu, follow the steps below:

STEP
To program the reading method, press:
METHOD:
READ TYPE: Endpoint
DELAY FIRST READ:
If incubator model: INCUBATION TEMP (Ambient or Temperature):
DELAY FIRST READ:

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140 | Appendix B: Programming a New Assay

STEP
WAVELENGTH (Single or Dual): Single
MEASURE (Wavelength[s] to use): 450
SHAKE (First, Every, or None):
To program the plate map, press:
MAP
AUTO
DOWN
DOWN
A01
BLANK MAP: AIR
NUMBER STDS: 05
NUMBER STD REPLICATES: 01
CONCENTRATIONS:
STD1: 6.25
STD2: 12.5
STD3: 25
STD4: 50
STD5: 100
REUSE STANDARD CURVE?
NUMBER CTLS: 02
CONTROL 1: PC
CONTROL 2: NC
NUMBER OF REPLICATES
PC: 01
NC: 01
SAMPLES: 89
SAMPLE REPLICATES: 01
To define a cutoff formula for Positive and Negative calls:
Note: Kit instructions specify that samples with concentration values greater than 11
should appear as positive. The ELx808 software calculates the cutoff based on
absorbance value or transformed value (see the previous example) and cannot calculate
based on concentration. The technician must make the positive or negative
determination visually, based on the calculated concentration results.
Curve: 4P
Note: As a general guideline, choose “linear” if you expect a straight line result. Choose
“4P” for all others, unless otherwise specified by the kit instructions. The reader will
automatically calculate the concentrations of the samples when the assay is run.

BioTek Instruments, Inc.

Appendix C

Adjusting the Power

Input Voltage Setting

This appendix does not apply to ELx808 readers equipped

with an external power supply.

This appendix provides instructions for adjusting the power input

voltage setting in ELx808 instruments that are equipped with an
internal four-voltage range power input module.

Identifying the Reader’s Power Input Module ...................... 142 

Newer-Style Module ........................................................ 143 
Adjust the Power Input Voltage Setting ......................... 143 
Reconfigure or Replace Fuses ....................................... 145 
Older-Style Module ......................................................... 147 
Adjust the Power Input Voltage Setting ......................... 147 
Reconfigure or Replace the Fuses ................................. 148 
142 | Appendix C: Adjusting the Power Input Voltage Setting

Identifying the Reader’s Power Input Module

Previous versions of the ELx808 (models manufactured before December 2005) have an
internal, four-voltage range power input module, instead of an external power supply.
Located on the right side of the ELx808, the input module can be adjusted for a 100, 120,
230, or 240 V~ voltage setting.
The voltage setting that is currently selected can be determined by observing which of the
four possible voltage indicator holes has a white peg in it. Check the peg now to make sure
the setting is appropriate for your location.
If you need to adjust the setting, first determine which type of power input module is
installed in the reader:
• The newer style module has a cover that can swing back toward the
reader, when opened with small needle-nose pliers. If your reader has this
module, follow the instructions starting on page 143 for adjusting the power
input voltage setting and reconfiguring/replacing the fuses.
• The older style module has a cover that can be detached from the reader
when opened with a small flat-blade screwdriver. If your reader has this
module, follow the instructions starting on page 147 for adjusting the power
input voltage setting and reconfiguring/replacing the fuses.

BioTek Instruments, Inc.

 Newer-Style Module | 143

Newer-Style Module

Adjust the Power Input Voltage Setting

Important! Turn off and unplug the reader before adjusting

the voltage setting.

1. Unplug the reader and remove the power cord.

2. Insert a small pair of needle-nose pliers into the two holes on the cover of
the power input module. Lift the cover open and swing it back toward the
power cord socket (see Figure 12 below).
3. As shown below, a small voltage selector card is located to the right of the fuse
holder. Using the pliers, carefully pull this card straight out of the
compartment.

Figure 12: Power input module: accessing the voltage selector card and fuse holder

ELx808 Operator’s Manual

144 | Appendix C: Adjusting the Power Input Voltage Setting

4. As shown in Figure 13 below, there are four different voltage range positions.
Rotate the white plastic indicator peg around the card so that it fits into the
correct groove for the desired voltage.

™ Point the peg upward so that the text matching the desired voltage
range is readable at the bottom of the card, as shown below.

White plastic
indicator peg
should point
upward.

This side (showing

text) should face the
ON/OFF switch
(voltage range
should be on the
bottom).

Figure 13: Voltage selector card input range positions

5. Reinsert the voltage selector card into the power input module so that the text
side faces the ON/OFF switch, and the desired voltage range is on the bottom
of the card (see above).
6. Reconfigure or replace fuses as needed (see next page).
7. Replace the cover, and verify that the white peg lines up with the correct
voltage indicator hole.

BioTek Instruments, Inc.

 Newer-Style Module | 145

Important! The peg indicating the intended voltage should

protrude through the cover. Do not power up the instrument
until the voltage range to be used is indicated correctly by
the peg.

Reconfigure or Replace Fuses

Important! Turn off and unplug the reader before

reconfiguring or replacing the fuses.

Either USA or International style fuses are installed in the fuse holder that is located
inside the power input module. The reader’s fuses are configured at the factory prior to
shipping. Use the procedure described below if you need to reconfigure or replace
fuses.

™ A failed fuse is usually an indication of another problem that a new fuse is

not likely to remedy. Contact BioTek’s Technical Assistance Center if the
fuse replacement fails to resolve the problem. See Chapter 1 for contact
information.

The fuse holder can have either one of two fuse configurations:
• 100/120 V (USA): A fused hot AG 1.5 A, 250 V Slo Blow (PN 46024), or
• 230/240 V (International): Both hot and neutral fused with a 5 x 20 mm, 0.63 A,
250 V Slo Blow (PN 46038).

To reconfigure fuses:
1. Turn off and unplug the reader.
2. Insert the needle-nose pliers into the two holes on the cover of the power input
module. Lift the cover open and swing it back toward the power cord socket.
3. Using the pliers, gently pull the fuse holder out of the power input module and
turn it over.

Changing to the • Remove the two 0.63 A, 250 V fuses.

100/120 V • Install one AG 1.5 A, 250 V fuse (PN 46024) on the
Configuration: fuse holder on the side opposite the original 0.63 A
fuse location.
• Reinstall the fuse holder with the fuse toward the
back of the power input module.

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146 | Appendix C: Adjusting the Power Input Voltage Setting

• Replace the cover, and verify that the white plastic

indicator peg lines up with the correct voltage
indicator hole.

Changing to the • Remove the single AG 1.5 A, 250 V fuse.

230/240 V • Install two 0.63 A, 250 V fuses (PN 46038) on the
Configuration: fuse holder on the side opposite the original AG 1.5
A fuse location.
• Reinstall the fuse holder with the fuses toward the
back of the power input module.
• Replace the cover, and verify that the white plastic
indicator peg lines up with the correct voltage
indicator hole.

To replace defective fuses:

1. Follow steps 1 through 3 on the previous page to access the fuse holder.
2. Remove the defective fuse and replace it with the correct new one.
3. Reinstall the fuse holder with the fuse toward the back of the power input
module.
4. Replace the cover, and verify that the white plastic indicator peg lines up with
the correct voltage indicator hole.

BioTek Instruments, Inc.

 Older-Style Module | 147

Older-Style Module

Adjust the Power Input Voltage Setting

Important! Turn off and unplug the reader before adjusting

the voltage setting.

Important! Be careful when removing the cover from the

power input module: the fuse holder is attached to the inside
of the cover.

Refer to Figure 14 on the next page for the following instructions:

1. Unplug the reader and remove the power cord.
2. Insert a small flat-blade screwdriver into the access hole between the power
cord inlet and the power input module cover. Carefully pry the cover off the
module. Note that the fuse holder is attached to the inside of the cover.
3. A small voltage selector card is located on the right side of the power input
module. Using a small pair of needle-nose pliers, carefully pull this card
straight out of the module.
4. Text printed on the card identifies the four different voltage positions. Each
voltage number has an arrow printed next to it. Rotate the white plastic
indicator peg so that it fits into the groove on the card that represents the
desired voltage; the peg should point in the opposite direction from the arrow
printed on the board.
5. Orient the card so that the peg is facing out and the text on the card is facing
the ON/OFF switch. The arrow should be pointing into the power input
module. Slide the card all the way back into the input module.
6. Reconfigure or replace fuses in the fuse holder, as needed (see the instructions
on page 148).
7. Replace the cover on the power input module and verify that the white peg
lines up with the correct voltage indicator hole.

Important! The peg indicating the intended voltage should

protrude through the cover. Do not power up the instrument
until the voltage range to be used is indicated correctly by
the peg.

ELx808 Operator’s Manual

148 | Appendix C: Adjusting the Power Input Voltage Setting

Figure 14: Power input module; accessing the voltage selector card and fuse holder

Reconfigure or Replace the Fuses

Important! Turn off and unplug the reader before

reconfiguring or replacing the fuses.

Both USA and International style fuses are installed in the fuse holder that is attached to
the inside of the power input module’s cover. The reader’s fuses are configured at the
factory prior to shipping. Use the procedure below if you need to reconfigure or
replace fuses.

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 Older-Style Module | 149

™ A failed fuse is usually an indication of another problem that a new fuse is

not likely to remedy. Contact BioTek’s Technical Assistance Center if the
fuse replacement fails to resolve the problem. See Chapter 1 for contact
information.

The fuse holder has two configurations:

• 100/120 V (USA): A fused hot AG 1.5 A, 250 V Slo Blow (PN 46024), and
• 230/240 V (International): Both hot and neutral fused with a 5 x 20 mm, 0.63 A,
250 V Slo Blow (PN 46038).

The fuse configuration (U.S. or International) that is currently being used by the reader
is determined by which fusing network is facing the inside of the power input module.

To reconfigure fuses:
1. Turn off and unplug the reader.
2. Insert a small flat-blade screwdriver into the access hole between the
power cord inlet and the power input module cover. Carefully pry the cover off
the module.
3. Use the screwdriver to remove the Phillips-head screw that anchors the fuse
holder to the inside of the cover. See Figure 14.
4. Turn the fuse holder over.
5. Reattach the fuse holder to the cover with the screw.
6. Replace the cover, and verify that the white plastic indicator peg lines up with
the correct voltage indicator hole.

To replace defective fuses:

1. Repeat steps 1 through 3 on the preceding page to access the fuse holder.
2. Remove the defective fuse and replace it with the correct new one. Do not turn
the fuse holder over, or you will change the configuration.
3. Reattach the fuse holder to the cover with the screw.
4. Replace the cover, and verify that the white plastic indicator peg lines up with
the correct voltage indicator hole.

ELx808 Operator’s Manual

150 | Appendix C: Adjusting the Power Input Voltage Setting

BioTek Instruments, Inc.