Leica DM R

Instructions
 Ernst-Leitz-Straße
D-35578 Wetzlar (Germany)
Leica Microsystems Wetzlar GmbH
Tel. +49 (0) 64 41-29 0

http://www.leica.com
Fax +49 (0) 64 41-29 25 99
© Leica Microsystems Wetzlar GmbH z Ernst-Leitz-Straße z D-35578 Wetzlar z Tel. (0 64 41) 29-0 z Fax (0 64 41) 29-25 99 Printed on chlorine-free bleached paper.
Order nos. of the editions in: English 933 789 z German 933 789 z French 933 789 z Part-No. 501-148 Printed in Germany IX/98/FX/K.H.
4th edition, issued in 1999 by:

Leica Microsystems Wetzlar GmbH

Ernst-Leitz-Straße
D-35578 Wetzlar (Germany)

Responsible for contents:

Marketing MQM, product management Tel. +49 (0) 64 41-29 25 19
Fax +49 (0) 64 41-29 22 55

2
Leica DM R
Instructions

3
4
Contents
Important notes on this manual ....................... 7 Incident light interference contrast ................ 99
Incident light polarization .................................. 100
Assembly and description Possible errors .................................................... 101
of components ..................................................... 8 Diapositive overlay device ................................ 102
Assembly/General information ......................... 8 Macro device ....................................................... 103
Light sources ....................................................... 10 Linear measurements ........................................ 106
Lamp change. ...................................................... 12 Thickness measurements ................................. 108
Lamphousings ...................................................... 16 TV microscopy ..................................................... 109
Filters and filter magazine ................................. 17
Specimen stages and condenser holder ....... 18 Care and maintenance ...................................... 111
Condensers (transmitted light) ......................... 21
Incident light components ................................ 26 Wearing and spare parts, tools ....................... 112
Polarizers/analysers ........................................... 32
Tube optics ........................................................... 35 Index ..................................................................... 113
Tubes ..................................................................... 37
EU-Conformity declaration ............................... 114
Diapositive overlay, macro device .................. 40
Eyepieces ............................................................. 42
Objective nosepiece and objectives ............... 45
Objective labelling .............................................. 47

Operation .............................................................. 53
Basic setting
for transmitted and incident light .................... 53
Filters ..................................................................... 56
Focusing, mechanical ........................................ 57
Basic functions of motor focus ........................ 58
Calibration of motor focus ................................. 62
Objectives ............................................................. 65
Tubes and eyepieces .......................................... 67
General specifications
Transmitted light illumination ........................... 68
Phase contrast .................................................... 73 Mains voltage: 100–115 V/230 V, ± 10%
Transmitted light darkfield ................................. 75 (E focus)
Transmitted light polarization ........................... 77 90 – 250 V (mech. focus)
Transmitted light interference contrast .......... 86 Frequency: 50 – 160 Hz ~
Incident light sources ........................................ 90 Power consumption: max. 160 W
Fluorescence ....................................................... 93 Use: indoors only
IGS and RC ........................................................... 94 Operating temperature: 10 – 36 °C
Incident light brightfield .................................... 95 Relative humidity: 0 – 80 % to 30 °C
Incident light darkfield ....................................... 98 Overvoltage category: II
Incident light oblique illumination ................... 98 Contamination class: 2

5
 17

16

15

12 11 10
9
13

14

7 2 1
6 5
4 3

Transmitted light path*

1 Light source (lamphousing not illustrated), 2 Filter magazine*, 4-pos., 3 Diffusing screen,
4 Aperture diaphragm, 5 Imaging system of aperture diaphragm, 6 Field diaphragm, 7 Polarizer*,
8 Condenser
Incident light path*
9 Light source (lamphousing not illustrated), 10 Filter magazine*, 4-pos.
Diaphragm module with:
11 Aperture diaphragm* or filter and diffusing screen, 12 Field diaphragm, 13 Reflector or filter cube
Imagine light path
14 Objective, 15 Tube optics/Bertrand lens*, 16 Tube, 17 Eyepiece

* not part for all outfits

6
Important notes on this manual
The Leica DM R microscope series consists of Special manuals are supplied with some addi-
several basic stands and a range of modular tional equipment such as photomicrography,
components allowing an almost unlimited microscope photometry (MPV), compensators,
variety of individual outfits. heating stages, interference attachments, etc.
Therefore this manual has been given a modular There are also extensive brochures on micro-
layout as well to show you other possible scopy, which can be ordered, as can extra
configurations besides your own. copies of this manual, from our agencies for a
The manual is divided into two main chapters: cover charge.
Assembly (including a brief description of each Numbers in the text, e.g. 1.2, refer to the illustra-
component) and tions, i.e. Fig. 1, pos. 2 in this example.
Operation.

Any alterations or additional information are

described on extra pages. There is a Attention:
supplementary manual for the automatic
version. The manuals are multilingual. Due to This manual is an integral part of the product
the spiral binding you can turn the language you and must be read carefully before switching
want to the front. The manual can be filed in the on and using the microscope! It contains
supplied folder with the transparent plastic important instructions and information for
tongues. safe operation and maintenance of the
product and must therefore be kept in a safe
place!

Text symbols and their meaning:

Special safety information is marked at the

edge by the lefthand symbol and highlighted
by a grey background.

Warning of hot surface.

Attention! This symbol means that incorrect

! operation can damage the microscope or its
accessories.

Explanatory note.

Item is not included in all variants of the micro-

* scope.

7
Assembly/General information
Unpacking
Please compare the delivery carefully with the Attention:
packing note, delivery note or invoice. We
strongly recommend that you keep a copy of these Fire hazard! Keep lamphousings at least
documents with the manual, so that you have 10 cm (4˝) away from inflammable objects
information on the time and scope of delivery such as curtains, wallpaper or books!
later when ordering more equipment or when
the microscope is serviced. Make sure that no
small parts are left in the packing material. Assembly tools
Some of our packing material has symbols You only need a few ordinary screwdrivers to
indicating environmental-friendly recycling. assemble your microscope. These are supplied
with the delivery. Replacements for lost tools
! Attention: can be obtained from us or from a tool shop
(Fig. 1), see list of spare parts on p. 112.
When taking the microscope out of its packing
and putting it onto the desk take care not to
damage the sensitive vibration-damping feet on
the bottom of the microscope.

Attention:

Do not connect the microscope and periph-

erals to the mains yet! (see page 53).

Installation site

! Attention:
Fig. 1 Assembly tools 5
Make sure that the workplace is free from oil 1 3 mm hexagonal 4
3
2
and chemical fumes. Vibrations, direct sunlight screwdriver 1
and major temperature deviations have a nega- 2 Crosstip screwdriver*
6
3 Adjustment key for
tive effect on measurements and photomicro-
Sénarmont compensator*
graphy. This and an ergonomically designed 4 Pol centering key (long
chair which can be adjusted in several positions version)*
are the basic prerequisites for fatigue-free micro- 5 Centering key (short
scopy. version)*
6 Allen key 2 mm (3 mm)*

* not part of all outfits

8
Setting the mains voltage
The instruments and accessories described
Microscopes with mechanical focusing (42.12) in the manual have been checked for safety
are automatically adapted to the local mains or possible risks. Before making any altera-
voltage in a range of 120 +-256 % / 230 +-206 % V. For tions to the equipment or combining it with
microscopes with motor focus (RE and RXE non-Leica components in a way not de-
models, Fig. 44), however, the selector switch at scribed in this manual, consult the Leica
the back of the microscope (2.6) must be set. agency for your region or the main factory in
Wetzlar! Any guarantee will be rendered in-
valid if the instrument is opened or modified
Attention: in any way by unauthorised persons or if the
instrument is used in another way than the
one described in these instructions!
For external power units the mains voltage
should always be set according to the sepa-
rate instructions supplied.

Electric safety
To ensure that the microscope and
accessories are in a perfectly safe condition,
please note the following advice and
warnings: The mains plug must only be
inserted into a grounded outlet. If an
extension cord is used, it must be grounded Fig. 2 Back of microscope stand
1 RS 232 C* interface, 2 Connection for 12 V 100 W transmitted
as well. Using the ground connection (2.4), light lamp* , 3 Connection for 12 V 100 W incident light lamp*,
any accessories connected to the 4 Ground connection, 5 Mains connection, 6 115/230 V**
microscope which have their own and/or a switchover, 7 Space for extra lamphousing or switchable
different power supply can be given the same mirror, 8 Fuses (T4A), 9 Lamphousing 106*: screw for opening
lamp housing 106, O o Not illustrated, on the top surface of the
ground conductor potential. Please consult
back of the microscope: plug connection* for photomicro
our servicing personnel if you intend to (lamp and shutter control)
connect units without a ground conductor.

1
3
2
4
5
8

6 7

9
Fuses Retrofitting additional light sources
When retrofitting the incident light illuminating
axis the microscope must be equipped with a
Attention: deviating mirror (3.1) with lamp mount. If you
want to use 2 light sources alternately in
The two fuses integrated in the mains transmitted and/or incident light, a switchable
connection (2.7: T4A, see spare parts list on deviating mirror (3.3, either manual or motor
page 112) come into action when the mains controlled) can also be retrofitted.
voltage selector is incorrectly set (motor The non-switchable mirror (3.1) is mounted to
focus only) or in case of internal electronic the left, the switchable mirror (3.3) from the
defects. For fuses for external power units back. To do this, remove the cover (using a
please see the relevant special instruction sharp object if necessary), or, if a mirror is
manual and spare parts list on page 112. In already in place, remove it by loosening the
the event of repeated fuse failure it is 4 screws.
important to consult our Technical service. Hold the mirror you want to fit on the
microscope with the flattened side of the lamp
mount pointing downwards. For switchable
Assembly of light sources
mirrors only: before tightening the screws hold
Up to 4 lamphousings can be adapted depending the mount for the switching rod (3.4) at an angle
on the microscope configuration. If only one light of about 45° to the longitudinal axis of the
source is used this is normally attached to the microscope. Remove the stopper from the hole
left side of the microscope. Only lamphousing 106 (22.4) or (61.7) with the 3 mm hexagonal screw-
(2.8) and the microflash (see separate instruc- driver (1.1).
tions) can be used for transmitted light).

Fig. 3 Deviating mirrors

1 non-switchable deviating mirror, 2 Lamp mount without*
mirror for second lamphousing, with clamp screw,
3 Switchable deviating mirror*, 4 Mount for switch rod,
5 Switch rod*

3
4

5
2

10
Insert the switch rod (3.5) into the hole and Lamphousing 106 z
screw into the mount (3.4). Screw the lamp
for 12 V 100 W halogen lamp and gas discharge
mount without the mirror (3.2) onto the left of the
lamps up to 100 W (Hg 50, Xe 75, Hg 100 W,
microscope.
spectral lamps). Like lamphousing 106, without
diffusing screen, but with centerable and
Motorized mirror only: first fix the holder with
focusable reflector and 4- or 6-lens collector.
the short screw in the top right drill hole, then fix
Quartz collector on request. Fig. 5 and 48.1.
the lamp mount with the 3 long screws.
Lamphousing 252
Tighten the 4 screws to fix the lamp mount(s).
for gas discharge lamps up to 250 W (Xe 50, Hg
Lamphousing 106 200 W), centerable lamp socket, focusable 4-lens
collector, focusable and centerable reflector. In
only for 12 V 100 W halogen lamp (centerable in x
preparation.
and y direction), focusable, two-lens collector.
Without reflector, with grooved diffusing screen,
Microflash
heat-absorbing filter, Fig. 2.8, Fig. 4 and Fig. 48.17.
for photography of fast-moving objects. Only in
Besides lamphousing 106, the following light connection with the electrically switchable
sources can be used for incident light: deviating mirror and a lamphousing (see special
instructions).

11
Spare lamps Lamphousing 106 z
See page 112 for code nos.

Lamphousing 106
! Important:
For incident light only (48.1)! Disassembled like
Disconnect from power supply (2.5), lamphousing 106 (see above).
disassemble using hexagonal screwdriver (1.1
and 3.2). Unscrew screw (2.9) and remove cover. 12 V 100 W halogen lamp
Move the collector to the front (48.19).
Disconnect from power supply (2.5).
Remove the defect lamp and put a new
Loosen screws (5.4 and 5.9) with crosstip
12 V 100 W halogen lamp into the lamp holder
screwdriver and flip up lid (5.1).
without tilting (4.1).
Pull cut-out plug slightly out of socket (5.11).
Unscrew screws (5.10) on the lamp holder and
! Attention: pull out the lamp holder (Fig. 6). Remove defect
lamp and insert new 12 V 100 W halogen lamp.
Leave the protective covering on the lamp until it
is in its holder. Avoid making fingerprints on the
lamp or wipe off immediately.
Close the lamphousing (2.9).
! Attention:
Leave the protective covering on the lamp until it
is in its holder!
Avoid making fingerprints or wipe off
immediately.

Fig. 4 Lamphousing 106*, opened

1 12 V 100 W halogen lamp in holder, 2 Collector, 3 Diffusing
screen

2
3

12
Lamphousing 106 z* Hg- and Xe lamps
! Attention:
Avoid switching on and off fre quently, as this can
Attention: impair the stability of the lamp and shorten its
life.
Hot Hg lamps cannot be reignited until they have
Danger: the following information is
cooled down. We recommend that you let new
extremely important and should be adhered
burners burn in for several hours without
to under all circumstances:
interruption if possible.
Always unp lug the power unit from the mains
It is a good idea to keep a record of the hours
before assembly work is carried out.
the lamp is in use and to compare with the
Wait for the lamphousing to cool down before
manufacturer’s specifications. Replace dis-
opening (at least 15 min.), danger of ex-
coloured, spent lamps.
plosion!
We cannot accept any liability for damage
Never touch glass parts of the burner with
resulting from a lamp explosion.
your hands. Remove any fingerprints or dust
carefully (perhaps using alcohol).
Adjust lamp s immediately after ignition (see
Attention:
page 90 ff.)

Always wear safety clothing (gloves and face

mask) when assembling Xe burners (danger
of explosion).

Fig. 5 Lamphousing 106 z*

1 Lid, flipped up, 2 Collector, 3 12 V 100 W halogen lamp with
holder 4, 9 Lid screws, 5 Reflector, 6, 8 x/y centering of
reflector, 7 Reflector focusing, 10 Screws for lamp socket,
11 Socket for cut-out plug Fig. 6 12 V 100 W lamp holder (LH 106 z only)

5
6
2
3 7

8
4 9

10 11 10

13
 ! Attention:
Undo the screws (5.10) on the lamp holder and re-
move the holder (Fig. 7). Remove the spent burner
Protect movable interior parts with foam rubber by loosening the clamp screws (7.1 and 7.3).
or similar in case of shipment.
To open lamphousing 106 z and 252: undo screws Insert burner as follows, adhering strictly to the
(5.4) and flip up the lid of the lamphousing. Pull above safety information:
the cut-out plug slightly out of the socket (6.11). Do not remove the protective covering yet (7.7).

Fig. 7 Lamp holders for gas discharge lamps*

1 Upper clamp, 2 Seal point of the burner, 3 Lower clamp,
4, 6 Drillholes for fixing the holder, 5 Sockets for cut-out plug,
7 Protective cover

Hg 50 Xe 75
1
1 7

2
3
3

5
6

Hg 100 Hg 100
1
Stab. 1

3
3

14
Lamphousing 106 z, Hg- and Xe lamps
! Attention:
Make sure that the lamp base and the power
Attention: unit have the same number. If the lamp base is
marked L1, for example, L1 must also be set on
Always insert burner so that the power unit to make full use of the lamp and
1. the lettering on the metal base is upright not to shorten its life.
after insertion (different diameters of the Move the collector to the front position with the
metal base for the Hg 100 and Xe 75 burners focusing knob (48.19).
ensure that these are always inserted the
right way up). If one of the bases is labelled
“Up”, it must therefore be assembled at the Attention:
top.
Remove the protective covering from the
2. If the lamp bulb has a seal point (7.2), turn burner (7.7).
the burner so that this point will be at the
side, not in the light path. Put the lamp holder with burner inserted into the
lamphousing and secure with the screws (8.9).
Apart from the halogen lamp the following gas Try moving the collector (48.19): it must not
discharge lamps can be used, all requiring dif- touch the power lead.
ferent lamp holders (Fig. 7) and power units:
Type Average life ! Attention:
Hg ultra high pressure lamp 50 W (alternating current) 100 h
Xe high pressure lamp 75 W (direct current, stabilized) 400 h
When closing the lamphousing make sure that
Hg ultra high pressure lamp 100 W (direct current, stabilized/non- stabilized) 200 h the pins of the cut-out plug engage in the
Hg ultra high pressure lamp 100 W (direct current, stabilized/
non-stabilized, type 103 W/2) 300 h
sockets (8.8). Retighten the screws of the lid.
Push the cut-out plug in as far as it will go.
Put the upper pin of the burner between the Attach the lamphousing to the microscope
clamps of the flexible power supply and clamp (page 16) and connect to the power unit
with screw (7.1). (compare mains voltage!).

Unscrew the stud (7.3) in the holder slightly,

insert the lower end of the metal base and
retighten the stud.

Exchanging the collector on lamphousing 106 z:

Move the collector to the rearmost position with
the focusing knob (48.19). Pull the focusing knob
of the collector outwards. The collector can
now be removed.

15
Lamphousing 106, 106 z Microflash
The microflash is assembled in the same way
! Attention: (only in conjunction with the switchable mirror
and a lamphousing).
Only lamhousing 106 (48.1) can be used for
transmitted light!
Ventilation
Remove the dust protection cover from the lamp
mount. Unscrew the clamp screw (3.2) with the
aid of the hexagonal screwdriver (1.1) so that the
screw on the inner surface of the lamp mount Attention:
does not protrude above the surface. Align the
lamphousing so that the screw engages in the Important: Make sure that the instrument has
corresponding indentation on the lamphousing. sufficient ventilation:
Tighten the screw to fix the lamphousing firmly Take care not to block the air supply
to the microscope. underneath the microscope and at the con-
nected lamphousings or the air vents on the
Filter mount top of the microscope with paper, etc. Fire
hazard! Minimum distance from inflammable
A filter mount (Fig. 9) taking up to four extra
objects 10 cm (4˝).
filters (50 mm diameter) can be assembled
between the microscope and the lamphousing
in the same way. When lamphousing 106 is used,
only 1 thick or 2 thin filters can be inserted.

Fig. 8 Lamphousing 106 z with Hg 50 burner

1 Lid, 2 Collector, 3 Burner (Hg 50), 4 Reflector, 5, 7 x/y
adjustment of the reflector, 6 Reflector focusing, 8 Sockets for
safety cut-out plug, 9 Lamp holder screws

5
2
3 6

7
9
9
8

16
Filter holder*/lamphousing Filter magazine*
Filters with a diameter of 50 mm can be inserted The best way to accommodate filters is there-
in the special filter holder (accessory, Fig. 9) fore in the filter magazine (Fig. 10, 42.8 and 42.15):
next to the lamphousing or in the microflash, or Loosen the 2 fixing screws to remove the filter
placed on the microscope base (27.3) in magazine. It is easier to remove if the four
transmitted light. controls are operated. Put the filters into the
slots (without holders!) and tighten the clamp
Microscope base* and condenser* screw. Always put the diffusing screen in the
position nearest the lamp . Put the label caps
Filters with a diameter of 32 mm and holders can
(10.3) onto the corresponding switch rods and
also be placed on the microscope base. The
align the lettering.
mount on the underneath of the condenser
The filter magazine is more easily replaced if all
holder (27.6) should only be used for the
4 filters are tilted to one side first by pressing the
polarizer or whole- or quarter-wave compensa-
buttons. Finally, check that all 4 filters can be
tors (57a,1 and 2).
switched in and out smoothly and tighten the
fixing screws. If thick filters get stuck, try putting
them in a different slot or altering their position
in the slot.
Filters situated between the microscope base
and the condenser may cause disturbing
reflections (this may be remedied by slightly
tilting the filter) and lead to strain birefringence
Interference filters must be inserted with the
in polarized light and ICT.
bright reflecting side towards the light source!

Fig. 10 Filter magazine T/R (for transmitted and incident light,

Fig. 9 Filter holder (intermediate unit), with lamphousing for Figs. 42.8 and 42.15), also available with only 1 pos.
max. 4 filters, dia. 50 mm (when lamphousing 106 is used, only 1 Filter holder (Ø 32 mm, non-mounted), 2 Clamp screw for
1 thick or 2 thin filters can be inserted) filter, 3 Switch rod with push-on label caps

17
Mechanical stages* no. 1187 and 1189 Stage no. 1086 U*
Size of stage plate 200 mm x 159 mm, movement with inverted stage bracket, for incident light
range of object guide 76 mm x 46 mm, with 0.1° only. Size: 160 x 150 mm, stage clearance: 123 mm.
verniers for registration of specimen coordi- Object guide no. 12* can be adapted.
nates. Removable specimen holder.
Rotary Pol stage*
Up to 110° stage rotation, clampable. Vertically
Precision stage on ball bearings, stage diameter
adjustable coaxial drive for specimen positioning.
179 mm, 360° scale division and 2 verniers
Maximum specimen weight 4 kg.
reading to 0.1°, 45° clickstops, can be activated in
any azimuth, Fig. 13. 3 M4 drill holes for attach-
Stage clearance 25 mm for fixed stage, 63 mm
ment of heating stages, object guide, etc.,
for interchangeable stage. 2 M4 drill holes for
Fig. 13.
attachment of heating stages.
Pol 3 adaptable object guide for specimen formats
The 1187 stage (Fig. 11) is especially designed
25 mm x 46 mm, 25 mm x 75 mm, 50 mm x 50 mm.
for transmitted light and fluorescence micro-
Interchangeable control knobs with clickstops
scopy, whereas the similar 1189 stage is for
at 0.1, 0.3, 0.5, and 2 mm object displacement in
incident light microscopy (i.e. for thicker and
x and y direction.
heavier samples; shorter coaxial drives and
sample holder without spring clip), but also for
Other stage variants are adaptable besides
transmitted light microscopy.
these standard models, e.g. the SCOPOSCAN
scanning stage.

18
Only for microscopes with fixed stage Only for microscopes with interchangeable stage
The stage is protected against transit damage Assemble the condenser holder* (12.10) first
by 2 foam blocks (Fig. 11). Push out the upper (see page 20). Loosen the stage clamp (12.1) and
block first. To remove the lower block, move the hold stage against the dovetail guide (12.4).
coarse drive* (42.12) slightly. The block can then Screwing the stage clamp only slightly, align the
be pushed out at the side. stage for specimens up to a thickness of about
1.3 mm (transmitted light specimens) so that the
! Attention:
top end of the dovetail guide is flush with the top
end of the stage clamp. For thicker specimens
If the microscope has a motor focus: (incident light) and heating stages the stage is
after switching on the microscope* (42.14) tip clamped lower down.
coarse focusing “Up” (44.2, page 58) 1 – 3 times
to make the stage move upwards slightly. The
foam block can then be removed at the side.
Keep the foam blocks in case the microscope
Then clamp the stage tightly, as otherwise it may
needs to be transported again, as long periods
tilt slightly when a heavy load is placed on it.
of vibration lead to damage!

Fig. 12 Assembly of condenser holder* and specimen stage*

1 Stage clamp, 2 Drill hole for clamping the condenser holder
(3 mm hexagonal screwdriver), 3 Condenser height adjust-
ment, 4 Dovetail guide, 5 Adjustable upper stop of condenser,
6 Stage rotation clamp (no. 1187 and 1189), 7 Universal
condenser with disc, 8 Centering screws for light rings/
IC prisms, 9 Lever for condenser top, 10 Condenser holder
Fig. 11 Transit protection for microscopes with fixed stage* (with slot for whole- and quarter-wave compensators)

6
1
7
2
3 8
4 9
10
5

19
Pol object guide* Condenser holder*
Move the object guide until the fixing screw can The microscope stage must be equipped with
be seen under the drill hole (13.1). Insert the the condenser holder (12.10) for transmitted light
object guide in the guide holes of the rotary work. The condenser holder enables various
stage and tighten the fixing screw with the condensers to be changed quickly and centered
hexagonal screwdriver. and takes components for polarized light
(Figs. 27.6 and 57.1). An adjustable upper stop
Attachable object guide* (12.5) guarantees a reproducible vertical setting
of the condenser (Koehler illumination).
The attachable object guide can be fixed on the
left, right or at the front (not illustrated) with the
Interchangeable stages only: to assemble the
two clamp screws.
condenser holder, either remove the stage or
move it as far upwards as possible. Loosen the
clamp screw (12.2) slightly with the 3 mm hex-
agonal screwdriver, slide the condenser holder
onto the guide pin and retighten the clamp
screw (12.2) (already assembled for fixed
mechanical stage).

Important! Do not mount at an angle, note the

stop!

Fig. 13 Rotary Pol stage* and Pol 3 object guide*

1 Drill hole for fixing screw, 2 Swing-in/out lever to hold
specimen slides of different formats, 3 Place to keep center-
ing keys, 4 Pairs of clickstop buttons, 5 45° clickstop, 6 Stage
rotation clamp

2
5

20
Survey condenser UCR and UCPR* universal condenser
Only in combination with the Bertrand lens and For objective magnifications from 1.6x (trans-
survey observation (without objective!) see p. 64. mitted light interference contrast ICT from
5x objective) with sledge changer, swing-in/out
UCE* universal condenser holder for condenser tops, coupled with 2
auxiliary lenses (14.2 and 14.5), i.e. homo-
For objective magnifications from 1.6x (trans-
geneous illumination of the specimen and
mitted light interference contrast ICT from 10x
Koehler illumination are guaranteed for all
objective) with sledge changer, swing-in/out
magnifications from 1.6x.
holder for condenser tops. When the condenser
top is swung out of the light path (objectives
1.6x – 6.3x) the field diaphragm takes over the
function of the aperture diaphragm (Fig. 14b).

Fig. 14 a/b UCR/UCE universal condensers

The UCPR condenser has the same construction as the UCR condenser
1 Condenser top, 2 Upper field lens, 3 Centering screw for light rings and IC prisms, 4 Fixing screw for condenser disc
(removed), 5 Lower hinged lens (field lens)

UCR UCE
1

2
4
4

5
3 3

21
Condenser discs* for contrast techniques Condenser tops*
for UCE, UCR, UCPR condersers
Both condensers can be fitted with discs for
various contrast techniques (HF = Brightfield, The following condenser tops are available
DF = darkfield, PH = phase contrast, ICT = trans- (Fig. 16):
mitted light interference contrast) (See Fig. 15). 0.90 S 1 Dry condenser top for glass spec-
imen slides up to about 1.2 mm.
5-position condenser turret for HF, DF, 3 PH po- For HF, DF (up to objective aper-
sitions (15.1). tures of 0.75), PH and ICT and
polarization contrast.
8-position condenser turret for HF, DF, 3 PH P 0.90 S 1 As 0.90 S1, but for polarizing
positions, 3 ICT positions, or HF, 3 PH positions, 4 microscopes.
ICT positions (15.2). Whole- and quarter-wave P 1.40 OIL S 1 for ultra high resolution in bright-
compensators (15.3 or 17.6) can also be used field and for polarized light
instead of ICT prisms for polarized light micro- (conoscopy) and for ICT; for glass
scopy. specimen slides up to about
1.2 mm.
Achr. 0.50/S 15 for intercept distances up to about
15 mm, e.g. for heating stages, for
BF and DF.

Fig. 15* Discs for UCR, UCPR and UCE condensers

1 5-position disc, complete, 2 8-position disc, position 3 not
yet inserted, cover plate (with label) removed, 3 Assembly
keys for light rings and ICT prisms, H = hole for brightfield,
PH = light ring for phase contrast, D = light ring for darkfield,
K = Condenser prism K for ICT, λ/4 = compensator for
polarization, X = holes for centering keys Fig. 16* Condenser tops for UC/UCE condensers

D PH H PH PH 1 PH 2 D H l/4 K

0.50/S15
P1.40 OIL S1
P0.90 S1

X 3 X 0.90 S1

22
Condenser top Light rings* and turrets*
Screw the condenser top (Fig. 16) onto con- For transmitted light darkfield (DF) and phase
denser (14.1). contrast (PH) the UCR, UCPR and UCE universal
condensers (Fig. 14) must be equipped with a 5-
! Attention:
or 8-position condenser disc (Fig. 15) with a set
of light rings DF, PH (17.7 and 17.8). Darkfield can
Move the stage as far upwards as possible with also produced with the special darkfield
the coarse drive (42.12 or 44.2). Move the con- condensers (Fig. 53). The 8-position disc with
denser holder downwards as far as the stop (12.3). ICT prisms K is required for transmitted light
interference contrast ICT.
Securing the condenser
The light rings are normally inserted into the
Align the condenser against the horizontal
disc at the factory, so that you can skip the
dovetail guide so that the two centering screws
following assembly instructions. You can tell
(12.8) point to the back towards the microscope;
that the light rings have been inserted by the
Flip the condenser top to the front (lever 12.9).
fact that the four annular stops can be seen in
Loosen the clamp screw (27.4) and carefully
the window when the inner plate is rotated and
push the condenser to the back as far as the
that the labels DF, 1, 2, 3 (17.3) appear in the
stop. Slightly tighten the clamp screw (27.4).
reading window.

Fig. 17 Fitting the turret plates

1 Upper cover plate with reading window, 2 Lower cover plate
(8-position disc only), 3 Label plates, 4 Turret (8-position in
illustration), 5 ICT prisms K for ICT interference contrast,
6 Quarter- (and/or whole-)wave compensator for polarized
light microscopy, 7 Light ring for darkfield, 8 Light ring for
phase contrast, 9 Adjustment screw(s)

1 2 3 4

5 6 7 8 9

23
Light rings* and discs* ● Fit ICT condenser prisms if used (see below).
● For 8-position turret only: Lay the cover plate
Remove the disc from the condenser after
(17.2) on the disc so that all drill holes
loosening the clamp screw (14.4). Take off the
coincide and fix with the 3 screws. Push the
cover plate (17.1) after unscrewing the 4 fixing
plastic labels (17.3) into the cover plate as
screws. For 8-position disc only: Also take off
follows:
the second cover plate (17.2) after unscrewing
● On the side opposite to the axis of rotation,
the 3 fixing screws.
corresponding to the light ring, i.e. O 2 for light

ring 2 S 1, O
D for darkfield, O
H for brightfield, etc.
Insert the light rings for phase contrast (17.8,
● So that the lettering is not upside down when
identified by the code nos. 1, 2, 3 and the
read, i.e. reading in a direction away from the
intercept distance S of the corresponding
outer edge of the turret.
condenser top, e.g. 2 S 1) into the small holes
● Label unoccupied positions with blank white
(Fig. 15/PH) of turret as follows:
plates if desired.
● Unscrew both centering screws (15.X) slightly
Screw the upper cover plate back on with the
with the supplied Allen key (15.3) so that the
4 screws and fix the disc back onto the con-
light rings can be inserted.
denser (14.4). Make sure that the disc can be
● When the light rings are inserted, their labels
rotated by 360°.
must be visible, i.e. pointing upwards.
● Keep to the order 1, 2, 3. Insert the large light
λ- and λ/4-compensator
ring for darkfield DF into the large hole (15.D,
with centering facility). The darkfield ring can Model for 8-position condenser disc (17.6):
only be inserted into 2 of the 4 large holes on Insert so that the notch engages in the spring
the 8-position turret. fin; fix with an Allen key (15.3).
● Using the Allen key, readjust the centering
screws until they do not protrude outside the
outer edge of the disc and the light rings
cannot fall out.

24
ICT condenser prisms* Mount the light rings for phase contrast and
darkfield if appropriate (see page 23). First lay
Remove the 8-position disc (15.2) by unscrewing
the round cover plate on the disc so that all drill
the fixing screw (14.4) (the 5-position disc is not
holes and windows coincide and then push in
suitable for ICT). Take off the upper and lower
the corresponding labels (17.3, e.g. 10/20 for 10x
cover plates after removing the 4 (3) fixing
and 20x objectives), as follows:
screws.
● On the opposite side (i.e. on the other side of
Insert the ICT condenser prisms K (17.5) into the the axis of rotation).
large holes (15.K) in the order of their code ● So that the lettering is not upside down when
numbers (i.e. K1, K2, K3). Insert the prisms so read, i.e. reading in a direction away from the
that the code, e.g. K1, is on the outside. Turn outer edge of the disc.
back the adjustment screw (15.X) if necessary,
turn back both adjustment screws in positions 3
and 4. Press the prism against the spring clip
and engage the catch on the underneath in the
guide groove. Tighten the left-hand adjustment ● Different labels may be necessary for
screw if necessary (the additional right-hand differrent objective classes (e.g. N PLAN,
adjustment screw in positions 3 and 4 is for PL FLUOTAR, HC PL FLUOTAR, PL APO), so
darkfield or phase contrast only and must always refer to the supplied optics chart for
therefore stay screwed back for ICT so that the prisms!
adjustment of the prism with the left screw is not ● Label unoccupied positions with blank white
obstructed. labels if desired.
● Carefully wipe any fingerprints or dust off the
prisms.

Replace both the cover plates with the 7 screws

and attach the whole disc to the condenser.
Mount the condenser top 0.90 S 1 or P 0.90 S 1 or
P 1.40 OIL S 1 (other condenser tops are not
suitable!).

25
Incident light reflectors*/ excitation filter, dichroic mirror and suppression
fluorescence filter systems* filter) or the incident light reflector or the
adjusting reflector (Fig. 18) into the turret (Fig. 20)
Remove the front cover of the microscope (Fig.
with the angled end of the dovetail guide first as
19) by strong pressure upwards at an angle.
far as the stop.
Insert the filter system (combination of

Fig. 18* Incident light reflectors* and filter systems*

1 45° BF reflector with neutral density filter* N, 2 DF darkfield
reflector, 3 Adjustment reflector (DM R series only), 4 Fluo- Fig. 19* Front plate with incident light turret
rescence filter system, 5 Bertrand lens module, 6 ICR module, Sticker with filter positions 1 – 4
7 POL system, 8 Smith reflector Stickers of corresponding filter systems or reflectors

4
3
2
1

8
N 7
6
5

Fig. 20* Incident light turret

1 Display of position in the light path, 2 Display of filter system
or reflector, 3 Marking of assembly position, 4 Filter system or
reflector or adjusting reflector Fig. 21* Slot-on neutral filter N for BF reflector

4 3 2 1

26
Up to 4 positions can be occupied by rotating Retrofitting the incident light axis*
the turret.
Microscopes that were not fitted with the
In combination with incident light darkfield, a
HC RF 4 IL* module at the factory can have it
neutral filter (Fig. 21) can be slotted onto the
retrofitted as follows:
BF reflector (for brightfield, polarized light and
The following components are necessary for
interference contrast) to avoid glare when
fluorescence (for IL-BF/DF/ICR additional
switching between illumination techniques.
components are required from the Technical
Service):
– HC RF 4 IL+ module, incl. 4 mm Allen screws
(22.2)
The adjusting reflector, Smith reflector and DF – Deviating mirror with mount for lamphousing
reflector can only be placed at opposite incl. 4 4 mm Allen screws (3.1) or switchable
positions. The 4 turret positions are each mirror (3.3)
marked on the left of the dovetail guide with the – Cover plate for the side of the stand (22.10)
numbers 1 – 4 (20.3). In addition the position – Lid for filter magazine mount, incl. 2 cross-
currently in the light path is indicated on the head screws (22.8) or filter magazine (Fig. 10)
outside of the turret (20.1). Self-adhesive labels – Ground glass disc for lamp centration in
indicating the positions 1 2 3 4 and the mount (22.5)
abbreviations for the filter blocks and the – Adjustment aid (22.9 or 18.2)
reflectors (e.g. D) are enclosed with the filter – Front cover with hole (22.12)
systems and reflectors. Stick the label 1 2 3 4 . – Diaphragm module (see p. 29 – 30)
in its place in the upper line on the front – 2 centering keys (1.5)
plate (Fig. 19). – Lamphousing 106 or 106 z, power unit(s) if
Then stick the labels with the abbreviations in required.
the corresponding fields underneath according
to the marking on the systems (20.2) and the
number indicated on the left on the filter wheel
(20.3). The Smith reflector (with two reflecting
surfaces and lenses, Fig. 18.4) and the DF
reflector (with ring mirror, Fig. 18.3) do not have
a label.

Push the front cover hard until it locks back into

place.

+
IL = incident light

27
Remove the front cover of the microscope
(22.12); it is no longer required. ! Caution:
Using the supplied 3 mm screwdriver unscrew Store upside down so as not to damage the
the 4 fixing screws (22.1) and remove the cover
optics. Protect from dust!
with built in tube optics from the microscope. Using the crosstip screwdriver, unscrew the 4
fixing screws (22.11) of the analyser mount and
remove it (this component will not be required
again as the analyser mount is integrated in the
HC RF 4 IL module, 22.6).
Using the 2 mm screwdriver, unscrew the 4 Al-
len screws on the lateral cover plate (22.10). This
plate is no longer needed. Please keep the Allen
screws.

Fig. 22* Retrofitting the incident light axis (only for BF, DF
and fluorescence! Pol and ICR components can only be
retrofitted by the Technical service)
1 Cover plate (tube optics) with 4 fixing screws, 2 RF 4 in-
cident light module with 4 fixing screws, 3 Lamp mount (with
or without reflector), 4 Mount for switch rod (for switchable
mirror only), 5 Ground glass screen for lamp centration,
6 Analyser mount, 7 3 control points for assembly, 8 Cover
plate or filter box, 9 Adjustment aid (reflector), 10 Lateral
cover plate with 4 fixing screws, 11 Analyser fixture (only
before conversion), 12 Front cover with hole

6
7
4 9
3 8
10
7
11
5 12
3

28
Push out the cover cap from the inside and clip Diaphragm modules
the holder with the ground glass screen (22.5)
The diaphragm module HC F has a centrable
for lamp centration in its opening in the stand.
aperture (23c.6 and 8) and field diaphragm
Insert the HC RF 4 IL module (22.2) into the stand
(23c.3 and 4), an engageable BG 38 red
from above, with the turret pointing to the front
attenuation filter (23c.11) and a switch for
and downwards, as follows: Holding the HC RF 4
blocking the incident light path (23c.12). Main
module in the longitudinal axis, tilt it slightly
application: fluorescence microscopy.
forwards. Carefully put the module into the
The diaphragm module HC RF has an additional
stand with the turret as high up in the front hole
decentrable aperture diaphragm for oblique
as possible.
illumination (23b.6 and 7); instead of the BG 38
Put four 4 mm Allen screws into the bore holes
filter and the light path blocking switch it has a
in the HC RF 4 module, move the module to the
light-blocking neutral density filter (23b.5),
right and to the front so that it pushes against
interchangeable diffusing screens (23b.9) and
the stops (22.7) and tighten the screws with the
an optional focusing graticule* (23b.10).
screwdriver.
Main applications: all incident light techniques
especially bright field and darkfield, polarized
! Attention: light and ICR reflected light interference
contrast.
Put the cover back on the microscope (caution:
There is also a special MPV diaphragm module
built-in optics!), align by moving to the front and
HC for microscope photometry, and the reflection
to the right (22.7) and secure with the Allen
contrast module HC RC (see separate manuals).
screws.
Fix the metal cover (22.10) to the side of the
Assembly of diaphragm module HC F*
stand with the 4 Allen screws (2 mm screw-
driver). Push into the slot (63.5) from the left as far as
Close the mount for the IL filter magazine possible.
with cover (22.8) and screw down the cover Functions → p. 93.
with 2 cross-head screws or attach the filter
magazine (Fig. 10).
Hold the front cover (22.12, with slit) against the
microscope and push slightly so that it clicks in
position.
Assembly of deviating mirror on page 10, lamp-
housing on page 16.

29
Assembly of diaphragm module HC RF* The diffusing screen set A (23b.9) can be turned
over and interchanged with set B. Turn the slit of
Insert the focusing graticule in the mount*
the screw (23b.1) so that it is horizontal. Insert
(23a/b.10), first slackening the clamp screw
the diaphragm module HC RF into the slot in the
(23a.10) if necessary, making sure that the
stand (65.9) as far as possible. Turn the screw
smooth side of the mount points inwards, the
slit (23b.1) to a vertical position; the diaphragm
rotatable mount with slit points outwards, see
module is now locked in position.
p. 64. Tighten the clamp screw only slightly.
Functions → p. 93 and 96.

Fig. 23 Diaphragm modules HC RF (a, b) and HC F (c)

1 Fastening screw, 2 Grip for pulling module out, 3 Field diaphragm, 4 Centering screws for field diaphragm, 5 Neutral density
filter N in/out, 6 Aperture diaphragm, 7 Decentration of aperture diaphragm, 8 Centering screws for aperture diaphragm,
9 Diffusing screen set A and B, 10 Focusing graticule with clamp screw, 11 BG 38 filter, 12 Interruption of light path, 13 Lever for
additional lens

a b c

10

1 9
2 2
3 3 13

45 6 7 8 11 12
10 4 6 8
HC RF HCRF HC F

1 2

Fig. 24 IC objective prism turret and slide

3
1 IC prism with code letter, 2 Stop pin, 3 Adhesive label with
5
code letters (for opposite position!), 4 Adjustment screw,
5 IC prism in slide (only ICR reflected light with Pol objective
nosepiece)

30
Objektive prisms* for interference contrast The turret is assembled in its mount to the
ICT/ICR objective nosepiece as follows*: Unscrew the
two fixing screws (25.2 and 25.3) on the
The prisms are already fitted into the turret at
underneath of the nosepiece with the 3 mm
the factory in various configurations. If you
hexagonal screwdriver, remove the cover plate
should want to change the prisms yourself:
(25.3), put the IC turret in position and press hard
make sure to push the prism mount against the
against the two stops (25.1). Fix in position with
guide pin (24.2) and do not screw the fixing
the two longer screws. It is practical to take
screws too tightly (use washers!) to avoid
interchangeable nosepieces off the microscope
strain. The code letters, e.g. A, must be visible,
for this conversion.
cf p. 48 and 86. Stick on adhesive label (24.3)
corresponding to lettering of opposite positions,
e.g. A.

* When the IC device is ordered as a complete outfit, these

components are generally assembled at the factory.

Fig. 25 Conversion of objective nosepiece Fig. 26 Objective centering nosepiece*:

1 Stop pins in objective nosepiece, 2 IC prism turret with 2 Screws for tube slit/IC objective prism turret changeover. The
fixing screws, 3 Cover plate other screws must not be loosened under any circumstances.

1 2 3

31
On the Pol centrable nosepiece (Fig. 26 and 38.2) Transmitted light polarizers*
the tube slit (compensator module, 38.6) must be
The polarizer for polarization contrast (27.3) can
removed instead of the cover plate. This is done
either be placed directly on the window in the
by unscrewing the 2 fixing screws on the top.
microscope base or inserted from the right into
surface (Fig. 26).
the mount on the underneath of the condenser
holder (27.6).
! Attention:
ICT/P polarizer (Fig. 28) only:
Imortant: Do not unscrew the other 4 fixing
Remove the black plastic cover ring (42.7) from
screws or the centration of the nosepiece axis
the microscope base by exerting strong
will be lost!
pressure.
Alternatively, single objective prisms in slides
(not illustrated) can be inserted into the
centrable objective nosepiece (54.13), but only
for incident light interference contrast ICR.

Fig. 27 Condenser and transmitted light polarization contrast* Fig. 28 ICT/P polarizer*
1, 5 Condenser centration, 2 Fixing screw for the turret plate, 1 Clamp screw for rotation, 2 Polarizer (at an angle), 3 Index
3 Polarizer (Ø 32 mm), 4, 5 Condenser clamp, 6 Mount for adjustment, 4 Index reading, 5 Lever for disengaging the
↔

whole- or quarter-wave compensator or polarizer (Ø 32 mm) polarizer, 6 Vibration direction of the polarizer , 7 Fixing
screw

1
4
1
5 2
2 6
6 3
7
4
5

32
Slightly unscrew the clamp screw (28.7) if ÷ parallel to the longitudinal axis of the mount:
necessary with the Allen key (1.5 or 1.4). Place for polarized light microscopic examinations
the transmitted light polarizer on the microscope with the analyser 360 (30.1). The analyser must
base with its straight outside edge parallel to be set at 90.0° at the crossed position (see page
the right outside edge of the microscope base. 77).
◊ vertical to the longitudinal axis of the mount:
When you notice the orientation slot click into this position is always used with analyser IC/P
position (left) retighten the clamp screw. (30.5) 45°, analyser 360 only. For ICR up to fov 20
only!
Reflected light polarizers*
Polarizer with whole-wave compensator
One of the following polarizers is used,
depending on the area of application. They are For qualitative reflected light polarization (29.2).
inserted as far as possible into the stand from The rotatable whole-wave compensator permits
the right (29 and 65.4) see also p. 99. extremely sensitive colour contrast, e.g. for
microscopy of anisotropic ores and metals such
! Attention:
as aluminium.

Hg and Xe lamps can destroy the polarizer, so Polarizer ICR

use protective filter (29.6)! With fixed vibration direction (N – S) (29.5), due
to built-in MgF2 plate up to fov 25, but not for
Polarizer R/P polarized light. For reflected light interference
For qualitative and quantitative reflected light contrast ICR the ICR reflector with polarizer,
polarization (29.1). The interchangeable Pol filter analyser and MgF2 plate can be used instead.
can be taken out and inserted in two positions:

Fig. 29a Reflected light polarizers*

1 Polarizer R/P (switchable vibration direction), 2 Polarizer
with whole-wave compensator, 3 Polarizer rotation, 4 Whole- Fig. 29b Protection filter* for Hg and Xe lamps in polarized
wave compensator rotation, 5 ICR polarizer light*

a b

1
2

3
4

33
POL filter system Analysers*
Reflector ICR
There are two different types of analyser for
The polarizer and analyser are in a fixed crossed reflected and transmitted light polarization and
position and combined with a 45° reflector. interference contrast techniques:
Inserted like filter systems and reflectors (see Assembly: remove cap and insert analyser from
p. 26). The ICR reflector has a built-in MgF2 plate the left (48.2 or 54.3) as far as possible.
as well: better homogeneity (fov 25) but not for
colour contrast. Polarizer and analyser are not Analyser IC/P
required in this case.
Polarization direction E – W, rotatable through
Protective filter approx. ± 7° (30.5). Combined with a whole-wave
compensator (λ) on its upper surface, so when
! Attention:
the analyser is inserted the other way up, red I
becomes active (30.7), see also colour chart on
When using Hg and Xe lamps, the polarizers p. 80.
must be protected by a special protective filter!
Analyser 360
Rotatable through 360° and reading to 0.1° (30.2),
vibration direction in 90° setting according to
DIN: N – S. Engageable (30.4) neutral density
filter in empty slot to prevent glare when the
analyser is switched off. A whole-wave
compensator is not integrated, so colour
contrasting is only possible for ICR reflected
Fig. 30 Analysers light interference contrast with a polarizer ICR
1 Analyser 360, 2 Precision scale with 0.1° vernier (clamp from the “DM L” range.
screw on the back), 3 Orientation scale (90° intervals), 4 Neu-
tral density filter switch, 5 Analyser IC/P, with whole-wave
compensator inactive, 6 Clamp screw and index, 7 Analyser
IC/P turned the other way round for use of whole-wave
compensator

2 34 5 6 6 7

34
Functional description Another important function of the tube lens is
correction of chromatic and other image
In all microscopes with infinite tube length (∞)
aberrations, such as astigmatism. This used to
the objective theoretically forms the image at
be performed by the eyepieces in former
infinity, which would be of no use to the
microscopes. Additional correction by the tube
microscopist.
lens, however, has proved to be far more
Therefore microscopes with infinite tube length
advantageous. Optimum colour correction
always need a tube lens that projects the
cannot be carried out by one single lens – a
intermediate image into the eyepiece. The
system of several lenses, some of them
magnification of an objective for tube length ∞
cemented, is used, so that it is more accurate to
thus depends not only on the focal length of the
speak of a tube lens system. The tube lens
objective, but also on the focal length of the tube
system is permanently integrated in the top
lens, which is 200 mm. The magnification of this
plane of the stand (22.1), designated as cover
system, i.e. objective + tube lens, is engraved on
plate in the instruction manual, except for the
the objective, while the tube factor is defined as
tube module HC L (→ p. 36). This module is
1x and therefore does not need to be engraved
available in interchangeable versions.
(according to DIN and ISO standard). Infinity
objectives that comply with these conditions are
Conversion of tube optics
identified by the code nos. beginning with the
figure 506. . . , 556. . ., 557. . . , 566. . ., 567. Remove the 4 fixing screws (22.1) using the hex-
Objectives for ∞ microscopes with conventional agonal screwdriver, remove the tube optics by
reference focal length fB = 250 mm can also be pulling upwards and mount the module of your
used, but the engraved magnification factor choice with extreme care.
must be corrected with the value 200 : 250 = 0.8x.
However, as the visible field is then enlarged by
the factor 1.25x, the edges of the image may be
! Attention:
blurred. The code nos. of these objectives for Make sure the components are completely
tube lens focal length 250 mm begin with 559. . . , clean – it is particularly important to check that
and 569. . .; an adapter (spacer ring 32/RMS or there is no dust or fingerprints on the
25/RMS is also necessary due to the RMS underneath of the tube lens. Screw in the four
objective thread (see Fig. 39). The mount fixing screws loosely, so that you are still able to
(labelled collar) may also require modification. move the module.

35
In the opened upper part of the stand there are 3 Tube optics HC P 1x/1.6x with Bertrand lense
stop points (22.7), with corresponding points in
With tube factor 1x, switchable to 1.6x,
the tube module and in the incident light module.
engagable focusable and centerable Bertrand
Carefully pull the tube module forwards and
lens. Iris diaphragm in intermediate image for
simultaneously to the right to ensure that there
isolation of small grains (15 µm for 100x
is precise fitting at these three points. Carefully
objective). Specially for polarized light micro-
tighten the 4 fixing screws.
scopy, but can also be used for all other
The following versions of the tube optics are
techniques (54.1, 54.2; 58), see p. 77.
available:
Integrated depolarizing quartz plate: prevents
the formation of interference colours due to
Tube optics HC E
polarization effects of tube prisms (pseudo-
With tube factor 1x dichroism) when the analyser is disengaged and
For brightfield, darkfield, interference contrast the polarizer engaged. Only effective with tube
ICT and ICR, polarization contrast, fluorescence. factor 1x, however. Not for spectral photometry.
An auxiliary telescope (51.1) with adapter (51.3) is When using tube factor 1.6x, remember that at
also required for phase contrast, but for this the high objective magnifications and apertures the
tube optics HC B (or HC V) with Bertrand lens is useful magnification (objective aperture x 1000)
recommended. may be exceeded, causing blurred images.
Quartz plate inactive.
Tube optics HC B with Bertrand lens
With tube factor 1x, engagable and focusable Tube module HC L 4/25
Bertrand lens. Without tube optics, only for adaption of HC L
Specially for the adjustment of darkfield, phase tubes from the DM L microscope range in which
and interference contrast and for survey the tube optics are integrated.
observation (p. 65) and observation of very fine
bores. For all other techniques, including
polarization contrast, but not for quantitative
polarization microscopy (42.2 and 50.2).

Tube optics HC V:
Magnification changer with Bertrand lens
With tube factors, 1x, 1.25x, 1.6x and focusable
Bertrand lens (adjustment DF, PH, ICT and for
survey observation), see p. 64.

36
Tubes (DM R series) P = This tube is also fully suitable for polarized light
microscopy, as the crosslines in the right-hand
A wide range of tubes for various applications is eyepiece are automatically aligned together with
available for the LEICA DM series of micro- the tube to the polarized light microscope.
scopes. E = Provision for lateral adaption of overlay device
(p. 40 and 101).
R = Back reflection of format outlines and measuring
The abbreviations in the names of the tubes spot possible for photomicrography and photo-
mean: metry.
25 = Eyepieces up to field of view index 25 can be used
HC = Tube system HC, only with HC PLAN and wide field (e.g. L PLAN 10x/25))
eyepieces, HC photo adapter components, HC Outer diameter of eyepieces: 30 mm
TV adapters. V = Variable viewing angle.
F = Phototube, i.e. apart from the binocular obser- L = DM L tube range with integrated tube optics.
vation part the tube also has a vertical photo exit
for adaption of photomicrographic equipment,
video cameras and microscope photometers.
B = Binocular tube, for visual observation only.
SA = Automatic focus compensation: if the binocular
viewing port set to the individual interpupillary
distance of the user (p. 67), changing optical path
length (which would cause a blurred image when
the magnification was changed and during photo-
graphy) is automatically compensated.

Fig. 31 Microscope tubes

1 BSA 25: binocular tube with focal compensation (shown with pair of eyepieces), 2 HC FSA 25 PR and HC FSA 25 P: binocular
phototubes with (PR) or without (P) back reflection, 3 FSA 25 PE: binocular phototube with provision for adaption of lateral
overlay device, 4 Switch rod for beamsplitter, 5 Mount for photo adapter, 6 Photo adapter clamp, 7 Clickstop for Pol eyepieces,
8 Socket for light trap control cable (PR tube only), 9 Connection for lateral overlay device, 10 Example from HC L tube range
with integrated tube optics (tube HC LVB 0/4/4)

1 4 2 5 6 4 3 5 6 10

7 8 9

37
BSA 25 HC FSA 25 PR
Binocular observation tube 25, Fig. 31.1 Binocular observation and phototube (31.2).
Viewing angle 30°, not for polarized light Like HC FSA 25 P, but with additional back
microscopy. reflection for the MPV microscope photometer.
Switchable light trap of the binocular port for
HC FSA 25 P microphotometry. Back reflection only at the
beamsplitter setting 50 % / 50 %.
Binocular obervation and photo tube (31.2).
Viewing angle 30°, also for Pol microscopes,
HC FSA 25 PE
with 3 clickstop positions of the beamsplitter in
the tube: Binocular observation and phototube (31.3).
Switch rod (31.4) Visual Photo Like FSA 25 P, but with additional provision for
|–––– 100 % 0% the overlay of transparent (diapositive overlay)
|–––––– 50 % 50 % or non transparent (macro device) masks, see
|––––––– 0% 100 % pages 40 and 102.

HC FSA 25 V Photo adapter tube HC FSA and HC L

Binocular observation and photo tube (31.10) Interchangeable photo adapter tube with
with variable viewing angle from 0 – 35° and vertical exit (32.2) or with vertical and horizon-
image erection, i.e. image of object appears the tal* exit (32.1) for all HC FSA tubes, with 2
right way up and the right way round. clickstop positions for switchable beamsplitter
2 switching positions: 100 % light to binocular (100% to the top exit or 100% to the back). The
port or 20 % visual and 80 % vertical. Not for photo adapter tube HC L* (not ill.) with fixed
polarizing microscopy. beamsplitter ratio 50 % / 50% is available as an
option for the HC L3T phototube (DM L series).

Fig. 32 Photo adapter tube for FSA HC tubes

1 Switchable photo adapter tube*, 2 Vertical photo adapter
tube, 3 Beamsplitter switch rod (not for HC L3T tube), 4 Clamp
screw

3 1 2 4

38
Assembly of photo adapter tubes Phototube Leica DM RD HC
Slightly loosen the clamp screw (42.1) on the Automatic microscope camera system with
side with the 3 mm screwdriver, remove black integrated observation tube and 0 – 35° variable
cover, place tube on microscope and align viewing angle, automatic focus compensation,
edges parallel to the microscope. Retighten overlay of measurement field and format
clamp screw (42.1). outlines, image erection; also for Pol eyepieces
(field of view index 28 for zoom setting 0.9x);
The supplied vertical photo adapter tube (32.2) zoom eyepiece system 0.9x to 2.5x for all exits,
can be used instead of the photo adapter tube motor-driven; external overlay facility; one addi-
with two exits (32.1) on any of the photo tubes. tional exit each for a second 35 mm camera and
This is attached by loosening the clamp screw a TV camera; intermediate image plane access
(31.6) with the 3 mm hexagonal screwdriver and for graticules in slide for documentation
then retightening. purposes; with control electronics (Fig. 33 and
special instructions).
Eyepiece adapter tube HC,
TV adapter HC
Photo eyepieces and HC TV adapters can be
inserted into the photo adapter tubes.

Make sure you are using the right combination,

depending on the type of eyepiece, photo
system (LD or MPS) and TV chip size!

Fig. 33 Leica DM RD HC phototube

39
Lateral overlay* Diapositive overlay device
The devices for diapositive overlay and The diapositive overlay device consists of the
macroscopy can only be adapted to the reflection optics, the illumination unit with 6 V / 4 W
HC FSA 25 PE tube (31.9) and Leica DM RD HC halogen lamp (34.8), the standard 5 x 5 cm slide
phototube (Fig. 33). frame (34.6) and the control for focusing the
transparencies. The halogen lamp is fed by a
These tubes have a side flange (31.9) to allow separate transformer.
attachment of the reflection optics (Fig. 34 and 35).
Assembly of the diapositive overlay device
The reflection optics are used for the
mechanical and optical adaption of the Align the reflection optics to the tube flange
diapositive overlay device and the macro dual (34.1) with the coupling ring (34.2) and fasten
zoom system. with screws. The guide pin must latch into the
groove of the mount. Screw the diapositive
overlay device onto the reflection optics with
! Attention: the coupling ring (34.2) in the same way. Again,
make sure the guide pin latches into position.
If reflection optics are not adapted to the
microscope (34a.1 and 35.3), an image cannot be
obtained. Multi-viewing attachment
This is attached between the tube and the
microscope (not illustrated). Max. fov 25, see
also separate manual.

Fig. 34a Diapositive overlay device on the HC FSA 25 PE

1 Tube flange, 2 Coupling ring for reflection optics,
3 Reflection optics, 4 Coupling ring for diapositive overlay
device, 5 Knurled ring for focusing, 6 5 x 5 cm slide frame,
7 Filter slot, 8 Illumination tube of lamphousings Fig. 34b Transformer

1 2 3 45 6 7 8

40
Changing the halogen lamp in the illumination Macroscopy device
Disconnect from power supply. This consists of the reflection optics (35.3), the
Screw out the Allen screw at the back and macro adapter (35.5) and the macrodual zoom.
remove the lamp unit from the lamphousing.
Take the lamp out of the socket and replace, Assembly of the macro device
making sure that the contact paths of the lamp
Screw the reflection optics (35.3) onto the tube
lie on the contacts in the socket.
flange with the coupling ring (35.2).
Do not touch the lamp bulb with your fingers due
Align the macro adapter (35.5) against the
to the danger of perspiration burning in.
macrodual zoom and secure with the threaded
ring (35.6).
After the lamp unit has been replaced in the
Fasten the macro adapter and the macrodual
lamphousing, the lamp holder can be adjusted
zoom to the reflection optics with the coupling
vertically by about 2 mm with the Allen screw
ring (35.4). Watch the guide pin.
from beneath.
Looking through the microscope eyepiece,
adjust the lamp to the height where the greatest
image brightness is achieved.

Fig. 35 Macro device on the HC FSA 25 PE tube

1 Tube flange, 2 Coupling ring, 3 Reflection optics, 4 Coupling
ring, 5 Macro adapter, 6 Threaded ring, 7 Zoom setting
ring 1 : 4, 8 Zoom factor scale, 9 Scale for magnification factor
of the working distance, 10 Scale for distance of object from
the lower edge of the mirror housing, 11 Mirror housing

12 34 5 6 7 8 9 10 11

41
For direct visual observation (see page 37 – 38 Eyepiece labelling
for tubes) only eyepieces of the type HC L PLAN
Example: 10 x/20 M (Fig. 36)
can be used. Fitting diameter = 30 mm.
This name is put together as follows:

10x

L PLAN type eyepieces may only be used on Magnification of the eyepiece, i.e. the magnified
microscopes of earlier series (= DM R label intermediate image produced by the objective is
on the right side of the microscope in black, additionally magnified by the eyepiece by the
not red!). engraved value (= eyepiece magnification).
PERIPLAN eyepieces, eyepieces from stereo-
microscopes or of manufacturers may not be Total magnification of the microscopes = Mob x Meye
used, as the full performance of the objectives (Reproduction scale of the objective x eyepiece
would then not be utilized. Exceptions to this are magnification)
the Leica/Wild 16x /14 B and 25x /9.5 B eye- Example: Objektive 25x/0.50, Eyepiece 10x/20
pieces, for which a special adapter ring is 25 x 10 = 250x total magnification
required, which is pushed onto the eyepiece
(37.2). If the tube factor is not 1x, the result must be
multiplied by tube factor as well. In the above
example, the total magnification after switching
to tube factor 1.6x would be 250x 1.6 = 400x.
Fig. 36 Eyepieces
1 – 4 Eyepieces ready for use by viewers without eyeglasses
(anti-glare protection 10 mounted or pulled up), 5 PHOTO
eyepiece, 6 10x/25M eyepiece disassembled, 6 Upper part,
7 Lower part, screwed off (applies also for 10x/22M, 2.5x/6M,
but not for 10x/20 and 10x/20M), 8a, b Retainer ring for
eyepiece graticules, can be screwed out, 9 Eyepiece
graticule*, 10 Anti-glare protection, removed for viewers
wearing eyeglasses (it can be pushed back with eyepieces Fig. 37 Widefield 16x/14 B eyepiece
10x/20 and 10x/22, insertable and remove pos. 8a or 8b). The 1 Clamp screw, 2 Spacer ring for Leica microscopes (must be
12.5x/6M model is basically the same as the 10x/25M eyepiece pushed upwards as far as the stop)

10x/20M 10x/25M
10x/20 10x/22M PHOTO
10 10

1
2
8b 3
4
6 8a 5 1
7
2
9 10

42
The tube factor is only engraved on the The field number of the eyepieces used must
microscope if it is not 1x. The HC P (Pol) tube correspond with the field performance of the
system has 2 switchable tube lenses, 1x and objectives. If the eyepieces have too high a field
1.6x, whereas HC V tube optics have 3 switchable performance for the field flattening of the
tube lenses. The Leica DM RD HC phototube objective, part of the field of view, e.g. the edge,
allows a continuous variation of the tube factor. may appear out of focus.

Useful magnification Objectiv series max. recommended

eyepiece field od fiew
The total magnification for visual observation
should not be more than 1000x the objective
15 20 22 25 28+)
aperture. In the above example (n.a. = 0.50) this
would be the case for a total magnification of Achromats
about 500x using tube factor 2x. C PLAN Achromats
N PLAN Planachromats
HC PL FLUOTAR® Semiapo.
When this threshold value is exceeded, e.g. with HC PL APO Planapochromats
100x/1.30 Oil objective, 10x eyepiece and tube
factor 1.6x the image may appear out of focus Object field diameter: If you divide the eyepiece
(empty magnification). field of view by the objective magnification, you
will get the real diameter of the observed object
/20, /22, /25 field. The eyepiece magnification is not part of
the calculation. For example, with the 10x/25
Field number (fov) of the eyepiece. The field
eyepiece and a 50 objective an object field of
number represents the diameter (in mm) of the
25 : 50 = 0.5 mm can be viewed.
intermediate image that can be viewed through
the eyepiece. This appears magnified by the
eyepiece factor. The microscope image in a 10x/
20 eyepiece therefore appears to be as large as
a circle of 200 mm diameter, observed from a
distance of 250 mm (250 mm = reference viewing
distance).

+)
Fov 28 at zoom factor 0.9 with photo system DM RD HC

43
If the tube factor (TF) is not 1x, this value must
be divided by the tube factor as well. Example:
The eyepiece can be used both with and without
Polarized light microscope or zoom system with
spectacles. When wearing spectacles, pull off
TF = 1.6x
or push back the anti-glare protection (36.7), as
Objectfield = 0.5 : 1.6 = 0.3 mm.
otherwise part of the field of view may not be
visible.
M
The eyepiece has a focusable eyelens (36.4) and Photoeyepieces*
therefore allows individual focusing of the edge
The HC L PLAN eyepieces (fitting diameter 30
of the field of view, inserted graticules or over-
mm) are designed for direct visual observation
laid markings. Adjustment range = ± 4 dioptres.*
only. Special eyepieces with fitting diameter of
The light-coloured ring (36.5) that becomes
27 mm and the engraving HC . . . PHOTO are used
visible under the adjustable mount marks the
for the adaption of photomicrographic equip-
setting for a person with normal or corrected
ment with a fixed magnification factor, e.g.
eyesight when used without a graticule (when a
DM LD and MPS systems and for special TV
graticule is inserted the standard setting is
adaption systems.
about 0.5 mm above this mark).
Assembly of eyepieces
Assembly of graticules* in M eyepieces
Only use identical eyepiece types (left-right)!
Important: Be extremely careful to avoid dust
Exception: polarized light microscopy: The right-
and fingermarks, as these will be visible in the
hand eyepiece on polarized light microscopes
field of view. The graticule diameter is always
has lines and a scale division (e.g. for length
26 mm for HC L PLAN eyepieces.
measurements, see page 105). Due to a double
clickstop (31.7) the right hand eyepiece can be
10x/25 and 2.5x/16 eyepieces only: Screw the
set with the crosslines aligned at the north
retainer ring of the underneath of the eyepiece
south/west position (horizontal/vertical) or at an
(36.6). 10x/22 and 10x/25 eyepieces only: Screw
angle of 45°. The crosslines then show the
out the bottom part of the eyepiece (36.8) and
transmission directions of the polarizers or the
screw out the retainer ring with a blunt blade.
vibration directions of the object in its brightest
Insert the graticule with the coated side
orientation (diagonal position).
downwards (in the direction of the objective) so
that any lettering is seen the right way round
when later observed in the viewing direction.
Screw the retainer ring and the bottom part of
the eyepiece back in.

* It is possible to extend the dioptre compensation by

having an ophthalmic optician center antireflection
coated spectacle lenses (2 – 3 dioptres) and inserting
them into the glare protection ring (36.7). However, this
method is not generally recommended by Leica.

44
Widefield 16x /14 and 25/9.5 eyepiece pair: push Objective thread and objective spacer rings*
the spacer ring (37.2) on to the lower part of the
Incident light bright- and darkfield objectives B
eyepiece as far as it will go secure with the
(40.1) have an M32 x 0.75 thread and can only be
clamp screw (37.1).
used on the objective nosepiece with M32
thread. These objectives have the letters BD
Objective nosepiece
after the aperture, e.g. HC PL FLUOTAR 10x/0.30
Depending on the type of microscope, the BD. Objectives with thread M25 x 0.75 can be
objective nosepiece is either fixed or inter- screwed onto all nosepieces. An adapter ring
changeable (Fig. 38 and 48.5). (M32/M25), Fig. 39, is available for using these
The following types of nosepiece are available: objectives on nosepiece BD with M32 thread.
Septuple objective nosepiece, M25 objective
thread, changeable and interchangeable
dto. coded, not interchangeable and inter-
changeable
Centerable sextuple Pol objective nosepiece,
interchangeable only
Sextuple objective nosepiece (BD), for incident
light bright-/darkfield
Objectives with M32 thread, interchangeable
and non-interchangeable
dto. coded, non-interchangeable and inter-
changeable

Fig. 38 Objective nosepieces

1 Septuple objective nosepiece (M25), 2 Sextuple centerable
objective nosepiece (M25) with tube slit and centering keys in
place, 3 Sextuple nosepiece (BD, M32), 4 plate, interchange-
able with IC turret (Fig. 25 – 26), 5 Objective centering keys in
place, 6 Tube slit, interchangeable with IC turret

1 2 3

4 5 6 5 4

45
Adaption of objectives with RMS thread (Royal Code numbers with a 6 or 7 in the third position,
Microscopical Society W 0.8x 1/36′′): objectives on the hand, indicate objectives for tube lens
with this classical thread size can only be used focal length 200 mm which is used without
on all nosepieces under certain circumstances exception in your microscope so that the en-
and together with the spacer ring M32/RMS or graved magnification applies.
M25/RMS (Fig. 39):
! Caution:
When using objective spacer rings:
Objective spacer rings are manufactured with a
Objectives with tube length 160 mm are not thickness tolerance of about 1/500 mm to ensure
adaptable at all due to optical reasons. These the parfocality of the objectives. They must
are identified by the engraving 160 and the mis- therefore be treated with extreme care. When
sing multiplication sign after the magnification, adapting objectives with RMS thread it may be
e.g. PL FLUOTAR 40/0.70. In the case of incident necessary to shorten the upper edge of the
light objectives whose engraved code number objective collar by about 1.5 mm (this is done at
has a 9 in the third position from the left, e.g. our factory) as otherwise the objective cannot
559 678 or 569 678, the engraved magnification be screwed on properly, so that parfocality is
must be multiplied by 0.8, as these objectives not guaranteed and the objective collar cannot
are designed for incident light microscopes with be rotated. Please consult our agency in this
tube lens focal length 250 mm. The aperture and case.
the working distance are not affected.

Fig. 39 Objective spacer rings (adapters)

25/RMS 32/RMS 32/25

46
Objectives/Assembly Lettering
For microscopes with fixed nosepiece: lower Example:
stage as far as possible (42.12 or 44.3). If you
have a motor focus, press keys 44.5 and 44.6 ∞/0.17/A N PLAN 10x / 0.25 PH 1 506 088
simultaneously to display an already stored
magnification (page 64).
∞
Microscopes with interchangeable nosepiece:
loosen the clamp screw on the left (48.5), pull Infinite mechanical tube length for which the
out the nosepiece towards the front and place objective is designed (there are also
upside down on a clean flat surface. microscopes and corresponding objectives with
Screw in the objectives carefully as far as tube length 160 mm), cf. Fig. 40 and 41.
possible in order of ascending magnification,
corresponding to the order of the light rings (PH 0.17
1 – 3) or the IC prisms in the condenser.
Stipulated specimen coverglass thickness. In
the case of dry objectives, the higher the
Once you have assembled the objectives and
aperture, the more important it is to keep to the
nosepiece, rotatable objective collars should be
cover-glass thickness of 170 µm. For an aperture
turned so that you can easily read the lettering.
0.85 the coverglass thickness should only
deviate a few µm at the most from 170 µm to

Fig. 40 Examples of objectives

1 Brightfield objective, 2, 3 POL objectives, 4 Phase contrast immersion objective, 5 Immersion objective with iris diaphragm,
6 CORR objective for inverted microscopes, 7 BD objective for incident light brightfield and darkfield (M25 thread)
Some immersion objectives with a knurled ring have a front part which can be pushed up and “locked” with a small rotational
movement. This device must be unlocked for observation! The sleeve of PL FLUOTAR and PL APO objectives can be rotated so
that the engraving can be read more easily.

1 2 3 4 5 6 7

47
achieve the full performance of the objective. A, B, C, D, E
We recommend coverglasses no. 1 H (high
Pupil position in the objective: the exit pupil of
performance, 0.17 – 00.02 mm) which comply with
most Leica microscope objectives has 4
DIN 58878/ISO 8255/1. The thickness of the
standard positions A, B, C and D, the so-called
embedding medium layer between the specimen
pupil blocks. When using the ICT and ICR
and the coverglass should be as thin as
interference contrast devices make sure that
possible. However, if you have a high dry
the IC prism (25.3 and 60.7) used above the
aperture and a non-standard coverglass
objective has the same letter, see “Optics” data
thickness, the aperture can be reduced by
sheet.
integrating an iris diaphragm (41.7) to make
deviating coverglass thicknesses uncritical.
The most important performance criteria of
Alternatively, an objective with correction
microscope objectives (apart from aperture and
mount (CORR) can be used.
magnification, see below) are field performance
and chromatic correction. Field performance is
0
understood as the diameter of the focused
Coverglass thickness 0, i.e. specimens must not intermediate image formed in the eyepiece (cf
be covered with a coverglass. These objectives page 43). As regards chromatic correction,
are primarily designed for reflected light there are three main types: achromats,
specimens, but can also be used to great semiapochromats (or fluorites) and apo-
advantage with transmitted light specimens chromats.
without a coverglass, e.g. blood smear
specimens. C PLAN
Achromatic objectives with a field performance
–
up to 20 mm (eyepiece fov max. 20).
The specimen can either be covered or not. A
maximum aperture of about 0.25 is considered N PLAN, PLAN
the threshold value for dry objectives for univer-
Planachromatic objectives with a field per-
sal use with or without a coverglass; for oil
formance of at least 20 – 22 mm. For visual
immersions this upper threshold is 1.25.
observation eyepieces with a field performance
of 20 or 22 mm are recommended, e.g. HC PLAN
10x/20. However, eyepieces up to 25 field of view
can be used if you are prepared to accept
slightly blurred edge definition.

48
PL FLUOTAR®, HC PL FLUOTAR, 0.25
HCX PL FLUOTAR
Numerical aperture of the objective, derived from
Semi-apochromats with a field performance of the angular aperture of the ray cone penetrating
at least 25 mm. The improvement in field the objective. The aperture influences a number
performance and colour correction compared of image factors and is therefore just as
with the achromats is particularly important for important as the magnification. It influences:
photomicrography. resolution, which also depends on the wave-
length λ of the light. A general rule for a medium
PL APO, HC PL APO, HCX PL APO wavelength λ = 0.55 µm for visible light is:
Plan apochromats with a field performance of λ = –––––
0.55
resolution = ––––
over 25 mm, the best objectives in the Leica 2 n.A 2 n.A
range.
Example: aperture 0.50
resolution (opt.) = 0.55 : 1.0 = 0.5 µm
PLAN L, N PLAN L
Achromats with particularly long free working Depth of field (axial resolution)
distances, specified in the Leica objective Image intensity: This increases quadratically
charts. L objectives with apertures over 0.25 are with the aperture, so objectives with high
designed for use without a coverglass. Field apertures, especially immersion objectives, are
performance over 20 mm. preferred for fluorescence microscopy, for
example.
PLAN H Coverglass sensitivity (cf 1st line 0.17!)
Achromats for use with heating stages which
have a 1.80 mm thick quartz window and with 1.25 – 0.65
interference attachments. Field performance Objective with built-in iris diaphragm to adjust
over 20 mm, the aperture (41.3), e.g. for darkfield immersion.
e.g. 10x/0.25 PH 1.

10x
Attention:
Magnification of the objective, which is also
indicated by colour of the lower edge of the
Objective with built-in diaphragm!
objective collar (see chart).
The knurled may only be used for adjusting
the diaphragm, not for screwing the objective
in or out.
Risk of damage!

49
PH 2 OIL
Phase contrast objective, with phase ring no. 2 Oil immersion objective: it may only be used with
built in. For phase contrast observation, the DIN/ISO standard optical immersion oil. For
corresponding light ring 2 in the condenser must apertures over 1.25 the engraving 0 or 0.17
be selected, see page 72. shows whether the objective should be used
Phase contrast objectives all have green with or without a coverglass. The coverglass
engraving. thickness should be adhered to as exactly as
possible (± 5 µm) for apertures larger than 1.32.
P Immersion objectives with an aperture greater
than 1.35 should only be used in a temperature
Extremely low-strain objective for polarized light
range of 20 – 25 °C. the refractive index of liquids
microscopy, with red engraving.
varies considerably at different temperatures,
the optical coordination between the objective
BD
and the oil changes during major temperature
Dry objective with M32 thread, for BF and DF fluctuations. The quality of the image may suffer
(incident light). in the same way as for the wrong coverglass
thickness. Also remember that if specimens are
↑ stained in strong colours, the temperature of the
immersion oil may rise by a few degrees due to
Leica objectives with infinite tube length can be
the object absorption. The illuminated object
used for both transmitted and incident light.
field should therefore be strictly limited to the
However, objectives corrected for coverglass
area observed (Koehler illumination, page 69)
thickness 0.17 are only used in transmitted light,
and the illumination intensity reduced if
as incident light specimens, of course, are never
necessary using a neutral density filter or the
covered (except for fluorescence specimens).
lamp supply.
The upwards arrow ↑ indicates that this
The immersion oil is applied with the stage
objective for use with or without a coverglass
lowered or the objective turned out of the light
should only be used in transmitted light, as
path, taking care to avoid air bubbles. It is later
disturbing reflections may occur in incident
removed with a clean cloth and ethyl alcohol, cf
light. This is indicated by the letter T instead ↑ of
p. 111.
arrow in the objective charts.
First read the safety data sheet (available on
request from your Leica agency).

50
W CORR Objectives
Water immersion objective. Use distilled, or at Special objectives with adjustable matching to
least demineralized water, if possible, as it is the coverglass thickness: Set correction mount
often difficult to remove the sediment from (not illustrated) approximately by turning the
drops of water that have dried on the objective. knurl to the average or estimated value: focus
the B specimen (→ Fig. 25).
IMM Adjust the correction mount until you achieve
optimum contrast, refocusing with the fine
Universal immersion objective for water, salt
control if necessary. This setting may be very
water, glycerine, oil.
difficult for specimens with low contrast or
weakly pronounced structures.
Locking of objectives
The front part (41.1 and 41.2) of certain Lens attachments
immersion objectives can be pushed in by about
Can be pushed onto the front of some objec-
2 mm and slightly rotated. This stops any
tives, or are readymounted at the factory:
remaining drops of immersion liquid from
wetting objects and other objectives when the
nosepiece is turned.

! Attention:
This locking device must be released before the
immersion objective is used again, as otherwise
the spring mechanism protecting the specimen
and the objective is inactive and the other
objectives are not parfocal with the immersion
objective. Fig. 41 Examples of immersion objectives
1 Immersion cap for N PLAN 10x objectives (pos. 2), 2 N PLAN
10x dry objective, 3 Achromat, 4 Planachromat, 5, 6 Objectives
with push-in locking device at front

1 2 3 4 5 6

51
Push-on cap CG and IMM Colour code rings on objectives
This can be used with some objectives with long In accordance with German and international
working distances to achieve optimum image standards (DIN/ISO) the magnification of each
quality with coverglasses (CG) of different objective is additionally indicated by a colour
thicknesses. Cap CG 0.4, for example, is ring above the knurl (41.4):
recommended for windows of vessels or for LCD 100x 63x 40x 25x 16x 10x 6.3x 4x 2.5x 1.6x 1x
displays with a thickness between approx. 0.25 125x
150x
50x 32x 20x 5x 1.25x

and 0.55 mm. Without CG cap 0.4 an optimum 160x

white dark light dark light yellow orange red brown grey black
image is achieved, for example, at a wall blue blue green green green

thickness of 0.95 to 1.25 mm (C PLAN L 40x/0.50

objective). Immersion cap IMM for enhancing Immersion objectives have a second coloured
contrast and observing inner reflections in ring further down (41.6):
incident light brightfield and POL (Fig. 41). black Oil or IMM
(= universal objective oil,
Reduction of reflections water, glycerine)
white water or IMM
A rotatable birefringent plate attached in front of
orange glycerine
the front lens can suppress reflections for
certain incident light objectives and thus
Engraving
improve image contrast. Used only with crossed
order code no. e.g. 506 001
polarizers or Pol filter system.
Six-digit factory code number of the objective.
Interference attachments Please always state this code number as well as
the full engraving of the objective when making
For quantitative measurement of roughness, film
technical or commercial enquiries. Objectives
thickness, etc. See special instruction manual.
whose code numbers begin with 569. . . and
559. . . can be used under certain conditions if
they have the engraving ∞, see page 45.
However, the engraved magnification value
must be multiplied by the correction factor 0.8x.
Objectives of tube length 160 or 170 (engraving
160 or 170) cannot be used at all.

52
Operation
Switching on Adjust brightness with dial (48.24). The numbers
are not absolute values, but merely enable
Turn on mains switch (42.14).
reproducible settings. The light-coloured dot on
Set selector switch to transmitted or incident
the dial indicates the setting for about 3200 K for
light (42.13). If using a gas discharge lamp: turn
photography on indoor colour film and TV
on external switch and check lamp adjustment
microscopy. See page 61 for DM RXE stand.
immediately (see page 90).
Tube optics
! Caution:
Disengage Bertrand lens (42.2), Switch on tube
Leica power units are immune to interference. factor 1x. If you have HC P (Pol) tube optics, just
Nevertheless we recommend you ignite gas switch to tube factor 1x (page 83). See page 67
discharge lamps before switching on the other for how to set tubes and eyepieces.
components, particularly if your power unit is
not from the Leica range.
Switchable mirrors (3.3, 61.7) only: Switch to left
or rear lamphousing.
Engage or disengage neutral density filter* (42.8,
42.15, 48.23, 65.10, 30.4, Fig. 9), depending on
Fig. 42
required brightness. 1 Tube clamp screw, 2 Bertrand lens* in/out, cf Fig. 50,
3 Reflector/filter system turret*, 4 Incident light polarizer*,
5 IC objective prism disc*, 6 Condenser disc*, 7 Coverring for
base of stand, 8 Filter magazine*, 9 Incident light diaphragm
module* cf Fig. 23, 10 Stage adaption*, 11 Place
to keep centering keys* (interchangeable stage only),
12 Mechanical coarse and fine focusing, 13 Transmitted/
incident light selector switch, 14 Mains switch with pilot
lamp* (not for motor focus), 15 Filter magazine* for
transmitted light

1
2
3 8
4
5 9

10
6 11
12
7 13
14
15

53
Analyser* Mechanical stages*
Disengage analyser (48.2) by pulling it out part Individual setting of specimen clamp:
way. Stage no. 1187: Push down the knurled ring (48.7)
on the joint of the specimen holder and turn to
Reflector*/filter system* the left (tighter clamping) or to the right (looser).
Then pull upwards so that it clicks into position.
For transmitted light only:
Disengage reflector (48.3) or filter system. Turn
Stage no. 1189: The clamping jaws can be moved
condenser disc (48.14) to pos. H (brightfield).
after the knurled screws have been loosened. In
addition a incident light object guide (code no.
For incident light only:
563 546) with movable sample platform for direct
Engage HF or Smith reflector (Fig. 18; 19; 48.3).
sample positioning and the tilting stage (code
For incident light fluorescence examinations of
no. 563 294) can be adapted.
transparent objects it is advisable to set
transmitted light mode first.
Individual setting of the x-y drive (Fig. 43):
Lengthening and shortening: First pull the lower
Adjustment specimen
control (for x adjustment, 43.2) downwards, then
For initial microscope adjustment we pull the upper control (for y adjustment, 43.1) in
recommend you use a specimen that has both the same direction.
high and low contrast areas. Non-plane parallel The coaxial drive is shortened by pushing the
reflected light specimens must be aligned on a controls upwards in the opposite order.
specimen slide with a handpress and plasticine.

Fig. 43 x-y specimen adjustment on the mechanical stage

1 y adjustment, 2 x adjustment, 3, 6 Clamp screws 4, 5 Rota-
table rings for torque setting

3 4
6 5

54
Torque setting: The torque has already been 0.3, 0.5, 1 and 2 mm. These are replaced by a
optimally adjusted at the factory, but you can strong axial pulling movement. Note the correct
change this setting as follows: move the lower orientation of the catch pins inside when
control (43.2) to the “long” position (see above). pushing on the new clickstop button. The stop
Push the upper control (43.1) upwards. screw on the underneath must be moved
Loosen the 1.5 mm Allen clamp screws (43.3 or inwards by about 2 mm to limit the vertical travel
43.6), using either an offset screw key (1.5 mm on smaller types of microscope.
socket-head) or one of the two centering keys
The two verniers permit angle measurements
(1.4 or 1.5). The threaded hole for the clamp
with a reading accuracy of 0.1.
screw of the upper ring is at an angle.
After 1 – 2 rotations of the rings (43.4 or 43.5) the
45° clickstop: Screw in the rotary knob (13.5) until
x and y adjustment can be set tighter or looser,
you feel slight resistance, then turn the stage to
respectively; move the x- and y-adjustment as
the next noticeable clickstop. Loosen the rotary
far as the stop if necessary. When you have set
knob, look for the position of the next clickstop
the torque, fix the ring with the clamp screw
(e.g. extinction position of object) and retighten
(43.3 or 43.6) and pull the upper control down.
the rotary knob. The stage can now be rotated at
clickstop intervals of 45°.
Stage rotation: Loosen the clamp screw (12.6).

Pol rotary stage*, Pol object guide*

The specimen is fixed to the stage either with
two spring clips or preferably, with the Pol 2
multi-format object guide (Fig. 13). For specimen
slides with a width of approx. 26 mm (1′′), swivel
out the metal plate (13.2) and insert the object as
shown in the illustration. If ordinary specimen
slides with a width of 26 mm are inserted
vertically to this, the movement range of the
object guide of about 30 x 40 mm is not fully
utilized. The supplied set of pairs of clickstop
buttons enables clickstops at intervals of 0.1,

55
Light filters* Green filter, Contrast enhancement for black-
panchromatic and -white photography.
Light filters can be built into the intermediate
filter holder (Fig. 9, filter diameter 50 mm), the
DLF 2 (blue) Conversion filters for colour
filter box (Fig. 10, Ø 32 mm) or can be placed on
photography with daylight film.
the dust protection glass of the microscope
base (27.3). Filters should not be used between
ALF dto. for artificial light film.
the polarizer and the specimen in polarized light
and ICT interference contrast (possibility of
BG 20 Highlights red in Polaroid expo-
birefringence due to strain caused by heat).
sures.
Besides the standard filters listed below there
are also various special filters, Optics data
VG 9 Contrast enhancement for chro-
sheet and interference filters for measurement
(green filter) mosome photographie.
purposes, e.g. the MPV microscope photometer.
546 nm Pol compensator measurements,
Filters Application interference interference attachments.
filter
Grey filter Grey filters (neutral density filters)
are used to attenuate light with- BG 38 Suppression of red in fluorescence
out influencing the colour tem- (blue filter) (is integrated in diaphragm mod-
perature. The engraved value, e.g. ule F (23.8).
N16, indicates the attenuation
value. N 16, therefore, means Diffusing For more homogeneous illumi-
reduction to 1/16 = 6.3 % trans- screen nation at objective magnification
mission. 1.6x and conoscopy and incident
Integrated grey filters can be light pupil illumination.
switched: in the microscope base
(48.23) (T = 6.3 %), Grooved Lamphousing 252 with 150 W
in the RF reflected light dia- diffusing Xe lamp.
phragm module (23.5), T = 5%, screen
in the empty slot of the analyser
360 (30.4) T = 25%,
Various grey filters can also be
inserted at the places described.

56
Stage clamp* Loosen clamp screw (48.9) on the left of the
stage bracket. Supporting the stage with both
Stage height setting (interchangeable stage
hands, carefully move it up or down.
only *)
The following chapters describe how to focus
the specimen. The stage height can also be ! Attention:
adjusted with the stage clamp (48.9). The stage Make sure the condenser does not touch the
should be clamped at the level where the microscope base.
thinnest specimens just touch the objective with Temporarily retighten the stage clamp.
the highest magnification at the highest possible Put the thinnest specimen you are going to
setting of the coarse/fine drive. As high-power examine (e.g. transmitted light object) on the
objectives always have telescopic front spring stage and move the stage up to the stop using
loading, there is hardly any risk of damaging the the coarse drive (42.12 and 44.2). Loosen the
specimen or microscope. stage clamp again (48.9) and carefully move the
stage upwards in the dovetail guide until the
! Attention: specimen just touches the objective with the
highest magnification, or an image can be
Don’t forget to release the locking mechanism focused.
on immersion objectives (page 51). If working with ordinary transmitted light
specimens of 1 – 1.2 mm thickness you can also
clamp the stage so that the stage bracket is
flush with the upper end of the dovetail guide
(12.4) after setting the upper stage stop.

Focusing, mechanical dual knob drive*

The smaller dial (42.12) is for fine focusing; one
division of the scale represents a vertical
movement of approx. 2 µm (see page 107). The
larger dial is for coarse focusing.

57
Motorized* focusing 1.2 Focusing
The position of the stage can be adjusted with
! Attention: – the focusing wheel (44.4) and
– the “Up” (44.2) and “Down” (44.3) keys.
Before using the motor focus, read the
On some models the interchangeable stage
instructions* carefully to eliminate the risk of
can also be vertically adjusted with the clamp
damage due to operation errors. If you have an
(48.9).
interchangeable stage, set the clamp (48.9) so
These controls are situated on both sides of
that specimens just touch the front lens of the
the microscope, giving you a choice of left-
higher-power objectives when the vertical
or right-handed operation.
adjustment of the stage is at its highest position.
1.2.1 Fine and coarse focusing with the
1.1 Switching on
focusing wheel
After you turn on the power supply with the
Like the mechanical coarse and fine focus, the
mains switch (42.14) the display (44.7) will still
motor focus also translates a rotary motion of
show the data set before the microscope was
the focusing wheel into a vertical motion of the
switched off last time, except for the “coarse
stage. One main difference, however, is that
drive” setting on the focusing wheel (44.4),
there is only one focusing wheel.
which is not stored.
Instead, the translation from the rotary to the
If neither the display nor one of the LEDs lights
vertical movement can be effected by keystroke
up, the microscope is probably not properly
(see below 1.3).
connected to the power supply (check mains
With the focusing wheel the stage can also be
cable connections).
moved over a set upper threshold (see section
1.4) but a lower threshold setting can only be
overridden by one step.
Fig. 44 Motor focus controls
Controls 1– 4 are situated on both sides of the stand in the
same layout.
1 Stepwidth, 2 “Up”, 3 “Down”, 4 Focusing wheel, 5 “Upper
threshold”, 6 “Lower threshold”, 7 Display

2 1

5
6

7
3 4

58
1.2.2 Stage height adjustment by keystroke indicated in the display (44.7). Each objective
position can be individually stored on the coded
The stage can be moved up and down at a
objective nosepiece, see page 60.
maximum speed of about 6 mm per second with
the “Up” (44.2) and “Down” (44.3) keys. At first,
The three possible stepwidth settings for the
the acceleration is deliberately retarded to
fine focusing are:
allow fine vertical movements by keystroke.
If the upper threshold has been set (see section 1 = 0.1 µm 2 = 0.7 µm 3 = 1.5 µm
1.4), the stage can be repositioned at this setting
with the “Up” key (with an accuracy of ± 1 µm). Coarse focusing
A set upper threshold cannot be overridden with
By simultaneously pressing the “Up” and
the “Up” key, but this can be done with the
“Down” keys (44.2 and 44.3) you can switch from
focusing wheel. If the z drive is above the upper
the set stepwidth to the “coarse drive of the
threshold, the stage will be lowered to the upper
focusing wheel” function. When the coarse
threshold when the “Up” key is pressed.
drive is activated, numbers 1 – 3 on the left-hand
side of the display light up simultaneously.
If no thresholds are set, the stage travels to the
With the coarse drive the stage can be moved
mechanical end-switch position.
up or down by about 1 mm per rotation of the
focusing wheel.
! Attention: The keystroke function of repositioning at set
thresholds is retained with full accuracy for the
Risk of damage, particularly to the condenser,
coarse drive.
the objectives and the specimens.
You can switch back to fine focusing by pressing
keys (44.2 and 44.3) simultaneously again.
1.3 Stepwidths, Focusing wheel
Fine focusing 1.4 Setting/deleting z thresholds
he motorized vertical movement of the stage is A threshold can be set at the current stage
not continuous, but by extremely fine repro- position by pressing and sustaining (≥ 1 sec) the
ducible steps. These are chosen, depending on “Upper threshold” (44.5) or “Lower threshold”
the objective, so that the stepwidth is smaller (44.6) keys.
than the depth of focus, giving the effect of
continuous focusing. The stepwidth for the
focusing wheel can be set with the “Stepwidth”
key (44.1). This alternates between three
possible settings when the key is pressed and is

59
You can delete a threshold whenever you like by The objective magnification and the offset to the
pressing the same key. focal plane must be “read in” once (see page 62,
The relevant key must be kept pressed down Calibration).
until the corresponding symbol in the display The stepwidth last used at a nosepiece position
field “z status” has switched over. The display is automatically stored.
then shows the active function: The setting of the stored stepwidth, the display
“Set ↑” setting of the upper threshold, of the magnification and the compensation of
“Del ↑” deleting of the upper threshold, the focus offset are done automatically while
“Set ↓” setting of the lower threshold, the nosepiece is rotated.
“Del ↓” deleting of the lower threshold.
If 2 coded, interchangeable nosepieces are
If you see the display “Err”! with flashing LED ↓ available, these can be labelled “nosepiece A”
or ↑ while you are trying to set a threshold, the and “nosepiece B” by operating a switch. As the
position of the threshold is not acceptable. system is capable of storing up to 14 objective
Examples: lower threshold = upper threshold positions, the data allocated to each objective
lower threshold > upper threshold. are automatically called up or displayed every
time. When you screw out an objective and turn
! Attention:
the nosepiece, you can see the switch inside the
nosepiece. This switch has to be switched to the
When viewing specimens of different thick- left for one nosepiece, and to the right for the
nesses, the upper threshold must be readjusted other (not illustrated). This can be done with a
every time the specimen is changed (risk of thin wooden stick or similar.
collision!).

1.5 Coded objective nosepiece*

The coded objective nosepiece enables several
parameters to be allocated and stored for each
objective position.
These parameters are:
– Stepwidth of the focusing (see section 1.3),
– Objective magnification (see page 64),
– Offset of objective focal plane (“parfocality”),
p. 62

60
1.6 Display 1.7 Collision and overload protection
Stage height
When an upper threshold is set, the height of the
! Attention:
stage in relation to the upper threshold is If the electronics register overload or a collision
indicated in the display, e.g. “-012”. while the motor focus is being operated with
The unit is displayed automatically with the two “Up” or “Down”, the motor is actively braked
LEDs µm and mm (i.e. 12 µm or 12 mm below the and switched off, and the display flashes.
upper threshold). Positive values signify stage In this case the stage should be immediately
positions above the upper threshold. moved clear in the opposite direction.
If the upper threshold is not set, you will see We cannot accept any liability for damage due
“Set?” in the display. If the lower threshold is to operation errors.
not set, the downwards arrow ↓ will not be
displayed. 1.8 Leica DM RXE microscope only:
Lamp voltage setting
Magnification display
With the exception of the Leica DM RXE
Regardless of the threshold status, you can microscope the lamp voltage is adjusted directly
switch between a display of the stage height with the dial (48.24).
and a display of the objective magnification (see
page 64) by simultaneously pressing the keys On the Leica DM RXE microscope, the dial
“Upper threshold” and “Lower threshold” (44.5 (48.24), which acts as a switch, must be slightly
and 44.6). turned clockwise until the voltage value of the
Switching over the display influences neither lamp (5 – 12 V) appears in the display (44.7); the
the thresholds nor the stage height. lamp voltage can be controlled with the
focusing wheel (44.4), if this position is
sustained.

When the switch is in the home position the

handwheel takes over the z drive control again.

This setting allows interactive adjustment of the

lamp voltage when a PC is connected. See
separate instructions for further details.

61
1.9 Parfocality However, it is not sufficient to focus on the edge
of the eyepiece field diaphragm or on diapositive
The depth of field (axial resolution) depends on
overlays (see page 101).
the objective aperture and the magnification; it
is under 1 µm for highest magnification
objectives. ! Attention:
In principle, it is possible to achieve absolutely
When the viewer changes his glasses or when a
perfect parfocality (identical focusing) of all
different person looks through the microscope
objectives used on the nosepiece by
the focusing of the eyelens(es) should always
mechanical and optical means, but this is
be checked and corrected if necessary. When
extremely complicated. It would be noticeably
the eyelens is not properly focused the focal
impaired even by the torque and any dust
plane of the objectives varies by different
particles on the objective shoulders when the
amounts, which can cause focusing errors and
objectives were screwed in. All the same, the
even collisions between specimen and
parfocality on Leica microscopes with
objective.
mechanical focusing is so precise that only
Adapted TV cameras may have a different focal
slight refocusing is necessary after each
plane compared with that for direct observation.
objective change. Using the motor focus, this
This may be caused by tolerances in the flange
parfocality can even be perfected with
focal length of the objective of the camera; the
automatic focus correction through the motor
flange focal length can be adjusted for some TV
focus and coded nosepiece for each objective
cameras.
after one calibration.
Objectives with coverglass information “0” must
not be used for covered specimens; only use
! Attention: objectives with the engraving “–” (i.e. for use
with or without a coverglass, see page 48) and
Please read the following important information
“0.17” (only with 0.170 mm coverglass). For
before storing the objective focus offsets:
heating stages with an observation window, H
Screw all objectives into the nosepiece with
PLAN heating stage objectives with engraving
about the same torque. If the nosepiece is
1.8 Q (i.e. for 1.8 mm quartz glass window) and
interchangeable, make sure it fits properly in the
“–” objectives can be combined.
microscope and keep the contacts clean. The
eyelenses of the eyepieces must be exactly
focused on the intermediate image. This is only
possible by inserting a (random) graticule in the
eyepiece or the Vario tube.
Another suitable focus indicator for the eye-
piece eyelenses is any overlay of a photomicro
device or the MPV microscope photometer.

62
Objectives with engraving “0” (i.e. without Accuracy can be enhanced by setting
coverglass) and “–” are suitable for uncovered variotubes and switchable tube lenses to a
specimens. If the microscope is used for both higher magnification factor or by putting the
covered and uncovered specimens, objectives auxiliary telescope (Fig. 51) on the eyepiece.
with the engraving “–” can be combined with Then set the upper threshold at this position
“0” as well as “0.17” objectives, without the with the key (44.5) (display 0 µm!) and switch off
focal plane having to be reprogrammed, with the the microscope (42.14).
exception of immersion objectives. Pressing key (44.5) at the same time, switch the
To store the focus data, always use a high- microscope on again. “OK!” appears in the
contrast specimen where the same area is display as long as the key (44.5) is pressed. After
suitable for all objective magnifications. For the key has been released “Cal!” appears in the
transmitted light the specimen used for storing display to indicate the storage of the focal plane
the focus data should be as thin as possible in of the first objective. “0” is now stored as offset
order to have a defined focal plane even at for the focused objective. Now all the objectives
highest magnifications, e.g. a Leica stage on the nosepiece can be focused.
micrometer.

1.10 Storing the objective focus offsets

Focus the specimen with the objective with the
highest resolution (i.e. max. aperture/magnifica-
tion).

! Attention:
When using immersion objectives (OIL, W, IMM):
release the locking mechanism of the front part
Fig. 44 Motorfocus controls
of the objective (page 51) to give the objective
Controls 1 – 4 are situated on both sides of the stand in the
the standard parfocalizing distance of 45 mm! same layout.
1 “Stepwidth”, 2 “Up”, 3 “Down”, 4 Focusing wheel,
5 “Upper threshold”, 6 “Lower threshold”, 7 Display

2 1

5
6

7
3 4

63
After focusing you only need to press key (44.5) Fit the survey condenser (cf Fig. 12, p. 23).
until “OK!” is output in the display to store the Remove objective or objective nosepiece.
offset. Finally, switch off the microscope briefly Focus the Bertrand lens* (50.3), open the
(42.14). aperture diaphragm = (48.21), the field
diaphragm (48.22) can now be used as aperture
1.11 Storage of objective magnifications diaphragm. For a more even illumination, a
diffusing screen can be used in the filter
As well as the offset values, the magnification of
magazine (42.15) or in the condenser holder B
each objective screwed in the nosepiece can be
(27.6). See also p. 80 and 102).
stored during the calibration. First store the
offset values of at least two objectives.
Incident light focusing graticule*
By pressing the key (44.6) and simultaneously
turning the focusing wheel you can set the Focusing can be made easier by inserting
magnification value of the objective currently in a graticule (23.11) into the diaphragm module
the light path. It is automatically stored when HC RF*, see page 29 – 30. After pulling out the
key (44.6) is released. diaphragm module part way (= channel II) this
During calibration it is not possible to set or graticule is projected onto the specimen surface
delete thresholds. To conclude the calibration and imaged together with it. This is particularly
the microscope must be temporarily switched useful for exact focusing of specimens lacking
off. in structures or contrast, e.g. for photo-
micrography or topological measurements.
Adjustment:
Attention! Only with Smith reflector! Set the
microscope exactly, particularly the eyepieces
The first time it is switched back on again the and the aperture diaphragm. Exactly focus a flat,
upper threshold for the focal plane (objective contrasty focusing object (e.g. incident light
with highest magnification only) must be deleted stage micrometer or mirror with scratches or
and reset. This also applies when the specimen other structures). Pull out the HC RF diaphragm
is replaced by a specimen of different thickness. module slightly = channel II, so that an image is
formed of the graticule.
Survey observation without an objective*
In transmitted light, the focusable Bertrand lens+
can also be used together with the survey
condenser (Fig. 45) as a survey objective with
ca. 1x magnification, making it possible to scan
objects with a diameter of about 25 mm (= width
of specimen slide). Not generally suitable for
photographic documentation. The DM RD HC
photomicro system can only be used from factor
1x, pronounced marginal fall-off (vignetting) is to
be expected.
+)
Max. SFZ = 25, not at tube optics HCP (Pol 1x/1.6x/Bert-
rand lens)

64
If this image is not absolutely sharp: remove the To immerse: Lower the stage or turn the
diaphragm module (see page 30) and slightly objective slightly out of the light path, apply 1 – 2
pull out or push in the mount of the graticule (slit drops of immersion oil to the specimen, taking
on one side for screwdriver). After replacing the care to avoid bubbles. Focus carefully, as the
module, check exact focusing and repeat the working distance of immersion objectives is
process if necessary! usually extremely short. Be careful with
objectives with front locking device!
Objectives
Centration
See page 47 for detailed information on how to
use objectives. The main points are described Only for polarized light microscopes:
again below: objective centration*
The objectives are centered by adjusting them
Objective engraving with two Allen keys (1.4) until the optical axis of
the objective (and thus the centre of the image)
Only use objectives with “infinite” tube length
coincides with the axis of rotation of the stage.
(∞ engraving).
When the objective is properly centred, a
focused area of the specimen does not drift out
∞ 0.17 0 –
of the field of view when the stage is rotated. A
Note coverglass specifications (objective en- specimen point in the centre of the crosslines
gravings 0.17, 0 or –). therefore remains in this position for a whole
stage rotation. It is advisable to use a high-
Immersions contrast specimen full of detail for objective
centration.
For all immersion objectives: before focusing,
make sure that the front part of objective is not
pushed in and locked (pull out telescopically,
page 51).
Only use OIL objectives with Leica DIN/ISO
standard immersion oil. Clean with ethyl alcohol
only.
Fig. 45 Survey condenser
IMM objectives can be used with water,
glycerine, oil, etc.
W objectives should be used with distilled
water.

65
Disengage the analyser (54.3), tube lens 1.6x Method II (Fig. 46b)
(54.11) and Bertrand lens (54.2). Greatly narrow Move the prominent point on the specimen (46a)
the aperture diaphragm (54.9). Insert the two to the centre of the crosslines M. Rotate the
objective centering keys above the objective stage until the point on the specimen is furthest
you want to centre (38.5). Focus the object. away from the centre of the crosslines M
There are two similar methods of objective (position A, Fig. 46b). Point A (= maximum
centration: distance of the specimen point from the centre)
may even be outside the field of view. Turning
Method I (Fig. 46a) the centering keys, adjust the image until the
Rotate the stage and note the point on the specimen point A is midway (= pos. B) between
specimen that remains stationary. This point pos. A and the centre of the crosslines M (46c).
corresponds to the mechanical axis of rotation Move point A to M and check that A stays at M
of the stage. when the stage is rotated (46d). Repeat the
Now move this prominent point of the specimen centering process if necessary.
to the centre of the crosslines with the two
centering keys. Rotate the stage and fine-adjust Each objective must be centered separately. If
the centration if necessary. an objective is screwed out of the nosepiece,
e.g. for cleaning, and screwed back in the same
place, its centration is more or less retained. If
the stage height is altered by a few centimetres
with the coarse drive or stage clamp (e.g. for
specimens of different thickness) the fine
centration may be slightly lost for all objectives.

Fig. 46a Fig. 46b

Centration method I Centration method II

M M M M
B B
A A A

a b c d

66
Tube and eyepiece setting Only if neither eyepiece has a graticule inserted:
When you adjust the eyelens a white line (36.5)
Set the beamsplitter in the phototube to the
becomes visible round the basic part of the
viewing position by fully or partly pushing in the
eyepiece. This indicates the correct position of
rod (31.4).
the eyelens for viewers with normal or
corrected eyesight.
The meaning of the switching positions is shown
Spectacle wearers must remove the glare
by symbols on the left face of the tube and
protection, but viewers not wearing spectacles
described on p. 38.
must always put it on (36.7).
Set the interpupillary distance by pulling apart
For eyepieces with graticule inserted only:
or pushing together (50.1) the eyepiece tubes
Defocus the specimen or remove from the light
until only one image can be seen with both eyes.
path and exactly focus the graticule by adjusting
Note your personal interpupillary distance,
the eyelens (Fig. 37.4) with a relaxed eye. (The
e.g. 65.
eye relaxes best if you look out the window
Close any tube exits (31.5, 31.9, 32 and 33) that are
at a far distant object for a moment). See also
not in use, as otherwise stray light can disturb
page 62. Only focus the specimen through the
the image.
eyepiece with graticule. Then close your eye
and focus the specimen by adjusting the second
eyepiece only.

67
Transmitted light lamphousing 106* Adjust the centering screw (48.18) for the hori-
zontal lamp adjustment with a screwdriver until
Remove any diffusing screen(s) and filters from
the blurred, bright, vertical line (= overlapping of
the light path (Fig. 9 and 10).
image and reflection of the filament) is in the
Method I:
centre of the bright circle. Reduce lamp
UCR and UCPR condenser (Fig. 14a):
brightness to do this if necessary.
turn in a 10x objective.
UCE condenser (Fig. 14b):
Adjust the centering screw (48.17) until the
turn in a 5x objective.
image of the filament is in the centre of the field
Raise the condenser to its highest position
in vertical direction as well (Fig. 47). Put the
(48.12).
eyepiece back on the tube and put the filters
Focus the specimen and find an empty area.
and diffusing screens back in the light path.
Switch the condenser disc (48.14) to position H
Alternatively, you can focus on the image of the
(= brightfield).
filament with a Bertrand lens or auxiliary
Disengage the condenser top (48.15).
telescope (Figs. 50 and 51, condenser top swung
Open the aperture diaphragm (48.21).
in, use objective 40 x to 63 x, swing out polarizer
Slightly narrow the field diaphragm (48.22).
48.25).
Remove one eyepiece from the tube and look
Method II:
into the open tube from a distance of a few cm.
Lay the adjustment device* (Fig. 47a) on the
Adjust the collector (48.19), looking through the
window in the microscope base and adjust the
eyepieces at the same time, until the reflected
image of the filament visible inside, as with
image of the lamp filament (Fig. 47a) can be seen.
method I, using the collector and centering
screws (48.19, 48.17, 48.18).

Fig. 47a Lamphousing 106

Reflection of the lamp filament, greatly schematized: in reality
the reflection is extremely low in contrast. In incident light the
bright overlap area is wider and less defined. Fig. 47b Adjustment device for transmitted light source

68
Brightfield, Koehler illumination Condensers, Field diaphragm
Setting of UCE, UCR, UCPR condensers and the Close the field diaphragm (48.22).
field diaphragm (Köhler illumination) Slightly narrow the aperture diaphragm (48.21).
Turn in a 10x objective or higher and focus the Swing in the condenser top (48.15).
specimen. Correct the upper stage stop for the E Turn the condenser stop screw (48.13) clockwise
focus (44.5) if necessary. The best position is and move the condenser to the top position with
just above the set focal plane. the height adjustment (48.12). Switch the disc
(48.14) to the H position (= brightfield). The disc
is not necessary for brightfield.

Fig. 48*
1* Lamphousing 106 z for reflected light, 2* Analyser,
3* Rotatable reflector turret, 4* Window for incident light
lamp adjustment, 5* Clamp screw for nosepiece change,
6* ∞ Turret for objective side Wollaston prisms, 7* Knurled
knob for adjusting the object holder, 8 Stage rotation clamp,
9* Stage clamp, 10 Centering keys for condenser disc,
11 Fixing screw for condenser holder, 12 Condenser height
adjustment, 13 Adjustable upper stop of condenser,
14 Condenser disc, 15 Lever for condenser top, 16 Condenser
centering screws (hidden, cf 27.1 and 27.5), 17, 18 Centering
screws for lamp holder, 19 Collector adjustment, 20 Focusing,
21 Aperture diaphragm, 22 Field diaphragm, 23 Grey (neutral
density) filter, 24 Illumination intensity control (12 V 100 W
lamp), 25* IC/P polarizer

1 2
3
4
5
6
9 7
10 8
11 14
12 15
13 16
17
18
The items marked with an asterisk 19
are not part of every outfit. 20 21 22 23 24 25

69
Turning the condenser stop screw (48.13) or the The field diaphragm (48.22) protects the image
condenser height adjustment (48.12), lower the from unnecessary heat and keeps all light not
condenser until the edge of the field diaphragm required for imaging away from the specimen so
is sharply focused (49b) and also centre the that contrast can be enhanced. It is therefore
image of the field diaphragm with the two only opened far enough to just illuminate the
centering keys (48.16 or 27.1 and 27.5) (49c). viewed or photographed object field. A change
of magnification thus always necessitates
Open the field diaphragm (48.22) until it just adjustment of the field diaphragm.
disappears from the field of view (49d). When
the objective is changed the condenser
centration may need slight correction. Adjust
the collector (48.19) until the image is
homogeneously illuminated.

Fig. 49 Koehler illumination

a Field diaphragm not focused, not centered, b Field
diaphragm focused but not centered, c Field diaphragm
focused and centered, but diameter too small, d Field
diaphragm diameter = object field diameter (Koehler
illumination)

a b

c d

70
Aperture diaphragm Replace the eyepiece or disengage the Bertrand
lens. For objectives with low contrast the
The aperture diaphragm (48.21) determines the
aperture diaphragm can be stopped down fur-
lateral resolution, depth of field and contrast of
ther to highlight faint specimen details. In
the microscope image. The best resolution is
polarized light microscopy narrowing the
obtained when the apertures of the objective
aperture diaphragm usually results in brighter
and the condenser are roughly the same.
colours except for conoscopy, see page 82.
When the aperture diaphragm is stopped down
to be smaller than the objective aperture,
resolving power is reduced, but the contrast is
enhanced. A noticeable reduction in the re- Attention: The aperture diaphragm in the
solving power is observed when the aperture illumination light path is not for setting the image
diaphragm is stopped down to less than 0.6x of brightness. Only the rotary brightness adjust-
the objective aperture and should be avoided ment knob or the neutral density filters should
where possible. be used for this.
The aperture diaphragm is set according to the An aperture diaphragm in the objective (41.3) is
viewer’s subjective impression of the image, the normally fully opened. The reduction in image
scale on the dial is just to allow reproducible brightness caused by stopping down results in:
settings and does not represent absolute Greater depth of field
aperture values. In principle you can do a Less coverglass sensitivity (p. 47)
calibration yourself by comparison with the Suitability for darkfield (p. 75)
apertures of various objectives. Visual Change in contrast
comparison of the apertures of the objective
and the condenser can be made as follows: Condenser top 0.90 S 1/P 0.90 S 1
Remove the eyepiece from the eyepiece tube or
engage an auxiliary telescope (Fig. 51) or Bert- The condenser top (48.15; 16) increases the
rand lens (50.2 or 54.2/54.11) and focus. Close or illumination aperture, which should be about
open the aperture diaphragm until its image is 0.6x – 1x of the aperture of the objective used.
just visible in the objective pupil (brighter circle). The condenser top may therefore only be swung
This is considered the standard setting, i.e. out for low-power objectives. The following rule
condenser aperture = objective aperture. of thumb applies for condenser tops 0.90 S 1 and
P 0.90 S1:

71
out/in P 1.40 OIL S 1
Objective magnification Condensor top S 1 The Condenser top P 1.40 OIL S 1 is used when
< 10x swung out maximum resolution is required with immersion
≥ 10x swung in objectives with an aperture > 1.0, or for polariza-
tion-optic conoscopy (page 82) of large shaft
angles. About drop of Leica immersion oil is
applied to the front lens of the condenser, taking
care to avoid air bubbles. The groove round the
For brightfield observation the condenser top
mount can pick up any superfluous oil.
can also be swung out for 10x objectives.
The oil condenser top and condenser top
However, DF, PH and ICT would not work with
0.50 S 15 are not intended for phase contrast and
the condenser top swung out.
ICT interference contrast.

When the condenser top is swung out, the UCR,

Diffusing screen, collector
UCPR and UCE condensers remain in the same
vertical position as when the condenser top is Image homogeneity can be optimized by
swung in. When the condenser top on the UCE adjusting the collector (48.19) and maybe
condenser is swung out, the field diaphragm engaging 1 – 2 diffusing screen(s) (Fig. 9 and 11).
takes over the job of the (variable) aperture
diaphragm. However, the “aperture diaphragm” Possible errors
must be fully opened with low magnifications Wrong coverglass thickness (see page 47) or
and the UCE condenser. There is no exact wrong objective. Specimen has been placed on
setting of the illuminated field. the stage with the coverglass downwards
With the UCR and UCPR condenser the field and instead of upwards.
aperture diaphragm functions are retained Aperture diaphragm (48.21) too wide or too
when the condenser top is swung out (Koehler narrow. Incorrect height or centration of
illumination). condenser.
Lamp not adjusted (page 68). Dirty optics.
0.50 S 15:
The Condenser top 0.50 S 15 is used from objective
magnification 5x. It has an intercept distance of
15 mm when there is no glass, etc. in the light
path between the condenser and the specimen.
The intercept distance is lengthened when plane-
parallel glass windows or liquids are introduced
into the light path by about a third of the
thickness of the glass or liquid, e.g. for a 3 mm
glass window the intercept distance is about
16 mm.

72
Phase contrast
Like transmitted light darkfield and transmitted
light interference contrast ICT, phase contrast is
used to produce high-contrast images of
unstained specimens.
Turn the phase contrast objective (engraving
PH) with the lowest magnification (generally
10x) into the light path and focus the specimen.
If you have trouble finding the specimen plane:
temporarily stop down the aperture diaphragm
(48.21) or use a stained specimen, setting the
condenser disc at H (48.14).

Set Koehler illumination (see also page 69):

sharply focus the field diaphragm together with
the specimen by adjusting the condenser in x, y
and z. Swing in the condenser top (48.15).

Set the light ring corresponding to the objective

engraving (e.g. light ring 1 for objective PH 1) on
the condenser disc (48.14).

Fig. 50 Tube and tube lens system with Bertrand lens

1 Interpupillary distance setting of the observation tube,
2 Dial for Bertrand lens (B) or tube lens (1x), 3 Focusing of Fig. 51 Auxiliary telescope
Bertrand lens 1 Adjustable eyelens, 2 Clamp ring for fixing the focus position

2
1

2
3

73
Open the aperture diaphragm (= pos. PH). Possible errors
Engage the built-in Bertrand lens* into the light
Specimen: too thick, too thin, too brightly
path by turning the knurled wheel (50.2) = pos. B,
stained; refractive index of mounting medium
and focus the annular structures (Fig. 52) with
and specimen identical so that there is no phase
the lever (50.3). See page 83 for how to operate
jump.
the Bertrand lens on the polarized light
Specimen slide too thick, so Koehler illumination
microscope.
not possible.
If your microscope does not have a Bertrand
Wedge-shaped coverglass position, so
lens: insert an auxiliary telescop e* (Fig. 51) into
centration of light and phase ring is no longer
the observation tube in place of an eyepiece.
effective.
Slightly loosen the clamp ring (51.2) and focus
Wrong light ring, or light ring has been put in the
the annular structures by adjusting the eyelens
turret upside down (see assembly on page 23).
(51.1). Retighten the clamp ring.
Aperture diaphragm not open. Wrong
condenser top (only 0.90 S 1).
Push in the two centering screws at the back of
the condenser (48.10 or 14.3) and rotate until the
dark ring (phase ring in the objective) coincides
with the slightly narrower bright ring (light ring
in condenser).

Disengage the Bertrand lens and watch the

quality of the phase contrast image. If using the
auxiliary telescope, watch the image with one
eye through the eyepiece. Then repeat the
centration process for the other objective light
ring combinations.
Fig. 52 Centration process for phase contrast, observed with
a Bertrand lens or auxiliary telescope
a Condenser in brightfield position (H), b Condenser in PH
position, light ring LR not centered, c Light ring and phase ring
centered

PH LR

74
Transmitted light darkfield Transmitted light darkfield
with UCE, UCR and UCPR condensers with special darkfield condenser
Darkfield is possible with all objectives from 10x Whether the DF condensers (Fig. 53) can be
magnification; the image background may be used depends on the aperture of the objectives.
inhomogeneously illuminated at lower magnifi- Objectives with built-in iris diaphragm (41.3)
cations. Solution for 5x objective: Use light ring 3 have adjustable apertures.
with condenser top swung out (UCR/UCPR
condenser only) or use condenser top 0.50 S 15 DF condenser: max. objective aperture
(condenser UCR/UCPR and UCE). The highest D 0.80 – 0.95 0.75
possible objective aperture is 0.75, although D 1.20 – 1.44 OIL 1.10
objectives with higher apertures can be used if it
is possible to reduce the aperture with a built-in Compared with brightfield objectives, phase
iris diaphragm. These objectives can be identi- contrast objectives do not produce such good
fied by the fact that the maximum and minimum imaging results for critical specimens in
apertures are given in the objective engraving darkfield.
and in our lists, e.g. 1.30 – 0.60, (Fig. 41.3).
Move the upper stop of the condenser to its
Rotate the condenser disc to position H (= bright- highest position by unscrewing screw (48.13) in
field). Focus the specimen (10x objective). If you clockwise direction. Put a specimen on the
have trouble finding the specimen plane, stage.
temporarily close the aperture diaphragm Carefully clean the upper and lower surface of
(48.21). Swing in condenser top 0.90 S 1. the specimen.
Traces of dust and oil film on the glass surfaces
Set Koehler illumination (page 69) (sharply focus or air bubbles in the mounting medium seriously
the centered field diaphragm together with the impair the quality of the darkfield image!
specimen). n.b.: Open the ap erture diap hragm (48.21) =
Open the aperture diaphragm as far as the stop pos. PH.
(= pos. PH) and turn the disc to pos. D (= dark-
field ring). Optimize image homogeneity by
slightly adjusting the height of the condenser
and collector (48.19).

75
Focus the specimen with the 10x objective, open Possible errors
the field diaphragm (48.22).
Darkfield illumination is very sensitive to the
Adjust the condenser in x, y and z direction
slightest inhomogeneities in the specimen. As
(48.12 and 48.16) until the field is homogeneously
dust particles and fingermarks on the upper or
illuminated, narrowing the field diaphragm
lower surface of the specimen and the front lens
(48.22). You can now switch to a higher-power
of the condenser also cause scattering and
objective. Make sure only the observed field of
diffraction of light, it is essential to keep
view is isolated by the field diaphragm.
specimen surfaces and neighbouring lenses
absolutely clean.
Immersion darkfield
If the objective aperture is larger than the
Assemble the immersion condenser (see above). threshold values listed above of 0.75 or 1.10, you
Before putting the specimen on the stage, apply will get an image similar to brightfield. This will
a drop of oil to the front of the condenser, also happen if the condenser is greatly
making sure there are no air bubbles. Set as for decentered.
“darkfield with UC/UCR condenser”, p. 75.

Fig. 53 Special darkfield condensers

1 D 0.80 – 0.95 (dry), 2 D 1.20 – 1.44 OIL, 3 Condenser bottom

1 2

3 3

76
Transmitted light polarization* Looking at the empty field of view, set the
optimum extinction position by rotating the
See page 65 for objective centration (polarized
polarizer (never the analyser!)
light microscopes only).
Adjust the light source, diaphragms and
condenser as for transmitted light brightfield
(page 69); the following description applies for
polarized light microscopes (Fig. 54) and for
other microscopes retrofitted with polarizers
(polarization contrast, Fig. 27).
Crossing the polarizers
Focus an empty area of the specimen or remove
the specimen from the light path.
Remove any compensators (50.13; 27.6), Bert-
rand lens (54.2 or 50.2) and incident light
reflectors (54.12) from the light path.
Rotate the condenser disc (54.16) to pos. H.
Rotate the objective nosepiece (60.7) to pos. H.
Insert the analyser (54.3) and preadjust as
follows, corresponding to the polarizer used:
Analyser IC/P (30.5) Polarizer Ø 32 mm (27.3)
Make index coincide insert from the right (27.6)
exactly, the λ mark must or place on the window Fig. 54 Controls on polarized light microscope
point downwards in the microscope base 1 Centration* of Bertrand lens, 2 Bertrand lens* on/off,
(27.3) or focusing, 3 Analyser, 4 Objective nosepiece clamp screw,
Polarizer IC/P (54.17) 5 Stage clamp, 6 Centration of PH light rings and ICT prisms,
IC setting = 90° 7 Condenser height adjustment, 8 Polarizer rotation clamp,
(i.e. vibration direction 9 Aperture diaphragm, 10 Field diaphragm, 11 Tube lens 1x/1.6x*,
12 Quadruple* turret for incident light techniques, 13 Com-
↔

N – S (54.17 )
pensator slot (tube slot), 14 45° clickstop (hidden), 15 Stage
Analyser 360 (30.1) Polarizer IC/P (54.17) rotation clamp, 16 Condenser disc, 17 Index adjustment of
Set exactly at 90.0° pos. 0° setting (vibration transmitted light polarizer
(DIN standard) direction E – W ↔)

1 11
2
3 12

4 13

14
15
5
6 16
7

8 17
9
10

77
Make sure the specimen, the condenser lenses Index adjustment on IC/P polarizer
and polarizers are clean, as this will affect the If the two index marks on the mount of the
accuracy of the setting. polarizer (28.4) do not exactly coincide when the
A particularly accurate method of setting this polarizers have been crossed: alter the index
position is to use the built-in Bertrand lens (54.3 adjustment with the centering keys (28.3 or
with 54.11) on the polarized light microscope as 54.17) until the index marks coincide. After this
follows: adjustment the crossed position of the
Turn a high-magnification objective into the light polarizers can be set reproducibly or checked.
path (e.g. 40x, 50x, 63x).
Open the aperture diaphragm (54.9) (pos. PH). Examinations in polarized transmitted light
Focus the Bertrand lens or auxiliary telescope
The following section is only intended as a
so that the slightly brighter circle in the centre of
rough guide to the various examination
the field of view is sharply defined. If you slightly
methods. Further details are to be found in the
adjust the polarizer you will see 2 dark stripes
Leica booklet “Polarized light microscopy”,
that close to form a cross when the polarizers
code no. 923 009, and in many books on the
are exactly crossed (55a). If objectives and
subject.
condensers without the engraving “P” are used,
the cross usually does not completely close.
Examinations
Only one polarizer
If you want to examine specimens with other
transmitted light techniques such as brightfield,
phase contrast and darkfield instead of with
crossed polarizers, it is usually sufficient to dis-
engage either the analyser or the polarizer.

Fig. 55 Crossing the polarizers, viewing with a Bertrand lens and a high-aperture objective
a exactly crossed, b not exactly crossed
Pos. a cannot be set at all if there is strain in the condenser or objective

a b

78
If the image is not bright enough, both the faintly (they remain dark when the polarizers are
polarizer and the analyser should be exactly crossed). It is not customary to examine
disengaged. A neutral density filter (30.4) can be specimens with the polarizers parallel, as this
used in the empty slot of the analyser 360 (30.1) method of identifying birefringence is not sensi-
to protect the viewer from glare when the tive enough.
analyser is disengaged. Coloured birefringent
specimens may exhibit differences in brightness Change in brightness when birefringent objects
and colour when the stage or polarizer is rotated are rotated
(when the analyser is disengaged). This When the stage is rotated, the brightness of
phenomenon is called dichroism or pleochroism birefringent (anisotropic) objects changes
and is an important indication for crystal exami- periodically. During a full rotation the object
nations. However, this effect can also be disappears four times after each 90° interval.
simulated on non-polarized light microscopes, The four dark positions are called extinction or
as these have no built-in depolarizing quartz normal positions. Exactly between each of these
plate, or if an incident light reflector has been extinction positions the object can be observed
left in the light path when transmitted light is with maximum light intensity. These are the four
switched. This also applies for the use of the diagonal or 45° p ositions. In the extinction
tubelens 1.6x on the polarized light microscope positions the object vibration directions run par-
(54.11). allel to the transmission directions of the polar-
izers, at maximum intensity the object vibration
directions represent the angle bisectors of the
polarizer directions. The crosslines in the (right-
hand) eyepiece of polarized light microscopes
Incident light reflectors or fluorescence filter can either be aligned at N – S/E – W, i.e. in the
cubes should disengaged during examinations polarizer directions, or at 45° angles, i.e.
in polarized transmitted light and transmitted light corresponding to the object vibration directions
interference contrast ICT. in the diagonal position.

Examinations
Crossed polarizers
The DIN and ISO standard vibration directions
are shown in the chart on page 77, but when the
polarizers are crossed the same polarization-
optic effects are observed when the polarizers
are transposed by 90°.
If the specimen contains many non-birefringent
or opaque particles, the analyser is frequently
turned out of the crossed position by a few
degrees so that these particles show up at least

79
Survey observation The analyser IC/P (30.5) has a whole-wave
Put a transmitted light specimen on the p olar- compensator on one side, which is activated by
izer. Swing in the condenser top and focus inserting the analyser the other way up.
through the condenser with a low-power When a compensator is engaged, the phase
objective, e.g. 5x. Even though this method does difference is increased or decreased (see
not claim to produce good imaging Fig. 56).
performance, it allows extremely fast scanning The vibration direction γ (i.e. corresponding to
of series of specimens, cf also macro device on the refractive index nγ with the greater
p. 102. refractive index) can be determined from the
colour changes. The quartz wedge (57.7) allows
λ/4 and λ compensator, Quartz wedge variable colour shifts on the polarized light
microscope.
Depending on the microscope model, the quarter-
and whole-wave compensators are either
Circular polarization
integrated under the condenser (27.6), or, in the
case of polarized light microscopes, in the 8- Only with polarized light microscopes in
position disc (17.6) (vibration direction λ is dia- transmitted light:
gonal ) inserted in the tube slot (54.13). The Birefringent objects exhibit four extinction
↔

tube slot is closed by a spring-loaded dust positions for one stage rotation. Particularly
protection flap. when scanning a large number of specimens,
some of the birefringent objects will always
happen to be at the extinction position. Circular
polarization is used for simultaneous obser-
vation of the interference colours of all objects:
Black Remove the specimen from the light path or find
Lavender gray an empty area of the specimen. Cross the
Gray blue
polarizers exactly – they must also be exactly at
1st Order

200
Yellowish-white
the N – S/E – W positions, i.e. the analyser must
–λ Vivid yellow
400 be set either at the 90° or 0° position (54.3).
Red-orange
Deep red –λ
–
4
600 Indigo
Sky blue λ
+–
4
Greenish blue
2nd Order

800 Light green

+λ
Phase difference

Pure yellow
1000 Orange red
Dark violet red
Indigo
1200
Greenish blue
3rd Order

Sea green
1400 Greenish yellow Fig. 56 Interference colours in relation to phase difference,
Flesh color or to thickness and colour change for the addition and
Crimson subtraction position of a whole-wave and a quarter-wave
1600 Matt purple compensator

80
Insert quarter-wave compensator (57.5) in the To perform the measurement, the compensator
tube slot. is introduced into the tube slot and adjusted
Push the quarter-wave compensator (57.1) into until the object to be measured is in its maximum
the slot underneath the condenser (27.6) and extinction position. For this purpose the object
rotate until the empty field of view appears at its has to be moved into a certain diagonal position.
darkest position (first cross polarizers exactly!). With HC P tube optics the measurement areas
can be isolated with an iris diaphragm (58-I with
Compensators for quantitative measurements 54.11) Further details are given in the
instructions for the use of the compensators.
Only in conjunction with polarized light micro- The following compensators are available:
scopes in transmitted light. Adjustable com-
pensators are used for exact measurements of Elliptical Brace-Koehler compensator (57.9)
phase differences. For a known specimen thick- Rotary compensator with compensator plate of
ness d and the measured phase difference about λ/10 difference. Measurement is carried
gamma (Γ) the birefringence ∆n‘ can be worked out in white or in monochromatic light.
out using the following formula: Measurement range up to approx. 50 nm.
d
Γ= d x ∆n’ [nm] or ∆n = –
Γ

Fig. 57a/b Compensators

1, 2 λ/4 and compensator for pos. 27.6. Only for polarized light microscopes: 3 λ/4 and λ compensator for 8-position disc (54.16),
4, 5 λ/4 and compensator for tube slot (54.13), 6 Rotatable λ/4 compensator (Sénarmont compensator), 7 Quartz wedge,
8 Tilting compensator, 9 Brace-Koehler compensator

81
Ellip tical Sénarmont compensator (57.6)) Tilting compensator K, measuring up to 30 orders
( λ/4 compensator in subparallel position) (57.7)
Measurement is executed in monochromatic For the measurement of phase differences in
light (546 nm), and a 360° rotatable analyser white or monochromatic light up to the
(30.1) is necessary. Normally this compensator is maximum phase difference mentioned above.
used to measure phase differences of up to the The compensator plate is made of calcite;
first order, although higher phase differences evaluation is based on simple calculation by
can be measured, too. However, the means of enclosed tables and the stated
compensator does not produce the entire phase calibration constant. A programmed computer
difference but only the amount that is in excess can be used for evaluation of measurements
of a whole wavelength or a multiple thereof. taken with tilting compensators. The necessary
Whole wavelengths must be determined with a formulae and parameters are given in:
tilting compensator, quartz wedge, or estimation Kornder, F. and W. J. Patzelt: The use of
of the interference colour. Accuracy is higher minicomputers to evaluate polarization-optic
than with the tilting compensator alone. compensator measurements. – Leitz Scientific
and Technical Information IX/1, 30 – 32, 1986.
Tilting compensator B (Berek compensator)
measuring up to 5 orders Conoscopy of crystals
Compensator (57.8) with MgF2 plate or
Only with the Leica DM RXP polarized light
measurements in monochromatic or white light
microscope: Birefringent crystals cause
of up to 5 orders phase difference. The phase
interference patterns (Fig. 59a/b) in the exit pupil
difference can be read directly from the sum of
of the objective (i.e. inside the objective). These
the two angles of compensation produced when
are also called conoscopic images. The shape
the compensator plate is tilted in both direc-
of these interference patterns and the way they
tions, from a supplied calibration chart.
change when compensators are used supply
information on the number of the crystal axes
(uniaxial or biaxial crystals), the orientation of
these axes and the plus or minus sign of the
birefringence (positive or negative birefringent
crystal).

82
As these interference patterns occur in the pupil Setting the microscope for conoscopy
they are not normally visible during normal
The most suitable object areas for conoscopy
microscopic observation (orthoscopy). Their
are those that show the lowest possible phase
observation can be improvised by removing one
differences (chart in Fig. 56).
of the eyepieces and looking into the tube with
Exact objective centration and exactly crossed
one eye from a distance of a few centimetres.
polarizers are essential for perfect conoscopical
Observation is better with the auxiliary
observation. Turn an objective with as high an
telescope for phase contrast (Fig. 51). Other
aperture as possible (e.g. 40x, 50x or 63x) into
crystals in the field of view disturb the
the light path. Open the aperture diaphragm
interference patterns of a crystal in the centre,
(54.9). Move the crystal you want to examine as
so that this needs to be isolated. This can only
near to the centre of the field of view as
be done with the polarized light microscope
possible.
(tube optics HC P, with Bertrand lens and iris
Turn in tube lens 1.6x.
diaphragm). This module also has a second tube
Narrow the iris diaphragm (Fig. 58, pos. I) to match
lens allowing additional magnification by a
the size of the crystal, stopping down the field
factor of 1.6x.
diaphragm (54.10) as well if necessary.
Push in the Bertrand lens (58B) and focus by
rotating the control until the interference image
or the circular bright-dark edge of the pupil is
focused. Centre the Bertrand lens if necessary.
This is done by inserting the hexagonal screw-
driver (1.1) into the two holes (54.1) in succession.
Align the right-hand eyepiece so that the
crosslines roughly correspond to the directions
of movement for the centration process.

Fig. 58 Functions of the der Pol tube optics HC P

Controls Orthoscopy 1x Orthoscopy 1.6x Conoscopy

Tube lense (54.11) 1x 1.6x 1.6x

Iris diaphragm doesn´t matter as matched of the field > object
of view
Bertrand lens not in light path out in
(54.2)
Polarizers in or out in or out crossed
(54.3 and 54.16) (not for dichroism/
pleochroism)

83
Adjust the collector (48.19) to an optimal setting, The optical character can usually be identified
using a diffusing screen (42.15) if necessary. even when only one of the optical axes is in the
viewing direction of the observer. In the
Determination of optical character orthoscopic beam the brightness of specimens
orientated in this way changes little if at all
Uniaxial crystals (Fig. 59a)
during rotation. In the conoscopic beam, only
Uniaxial crystals observed in the conoscopic
one of the two isogyres will then be visible.
(divergent) beam show a dark cross, whose
centre indicates the position of the optical axis.
Biaxial crystals (Fig. 59b)
The cross is surrounded by coloured inter-
For the determination of the optical character
ference fringes*. When a variable compensator
cutting directions are particularly suitable in
(quartz wedge or tilting compensator) is oper-
which the bisectrix of the two optical axes is
ated the rings drift towards the centre or out-
parallel to the viewing direction (section vertical
wards in two opposite quadrants of the cross.
to the acute bisectrix).
The optical character is determined from the
In the divergent beam a dark cross will be seen
direction of movement of the rings as in Fig. 59.
which opens up into the two branches of a
Cutting directions in which the optical axis of
hyperbola, the so-called isogyres, when the
the crystal is inclined to the direction of
stage is being rotated. The cross and the
observation are suitable for the determination of
branches of the hyperbola are surrounded by
the optical character, which can mostly be
interference fringes. According to Fig. 59 or the
determined even when the centre of the cross is
rule mentioned below the optical character can
outside the field of view. Fig. 59 shows that fixed
be determined from the displacement direction
instead of variable compensators can also be
of these fringes after operation of the com-
used for the determination of the optical
pensator. The symmetry plane of the isogyres
character.
(axial plane) must be vertical to the γ direction of
the compensator.

* Only the cross is visible for thin specimens or

specimens with low birefringence.

84
Biaxially positive crystals:
The interference fringes move from the convex
to the concave side of the isogyres when the
compensator is operated.

Biaxially negative crystals:

The interference fringes move from the concave
to the convex side.

Possible errors
Polarizers damaged (discoloured) by powerful
light sources or dirty.
Objectives or condenser strained through
mechanical damage.
Beamsplitter or filter between the polarizers.
Mounting medium for transmitted light
specimens is birefringent.
Further sources of error on page 72.

Fig. 59a Determination of the optical character of uniaxial structures

Left: Positively uniaxial crystal, cut vertically to the optical axis
Right: Negatively uniaxial crystal, cut vertically to the optical axis
1 Diagram of the vibration directions in the object and in the compensator
2 Change in the interference pattern when a quarter-wave compensator is used
3 Change in the interference pattern when a whole-wave compensator is used

Fig. 59b Chart for determination of the optical character

85
Condenser Push the analyser (50.1) into the microscope as
far as the second clickstop. The lambda sign (λ)
Important: Only use the condenser tops
must be on the underneath (not visible), see also
0.90 S 1 or P 0.90 S 1 and P 1.40 OIL. The con-
page 34.
denser top with long working distance 0.50 S 15
(p. 22) is not designed for ICT work. See pages
Loosen the analyser clamp (30.6) and adjust so
25 and 30 for assembly.
that the two index marks are exactly opposite
each other. If using the 360° rotatable analyser
Crossing the polarizers
(30.1), set the 0° position with the coarse and
fine scale and vernier (30.2 and 30.3), clamp on
back. Retighten clamp.

To obtain a good quality ICT image the analyser Turn the objective nosepiece turret (60.7) and
must be set exactly at 0 and the polarizer must the condenser disc (60.8) to pos. H = brightfield.
be exactly crossed (extinction position)! The IC prisms are then disengaged. Disengage
the incident light reflector (60.6).

Fig. 60 Controls for ICT transmitted light interference contrast

1 Analyser, cf Fig. 30, 2 Stage rotation clamp screw,
3 Condenser top lever, 4 Polarizer rotation clamp, 5 Pola-
rizer index adjustment (cf Fig. 28), 6 Incident light reflector
turret, 7 Objective-side prism turret with fine adjustment,
8 8-position disc for condenser-side Wollaston prisms,
9 Mount for λ or λ/4 compensator (hidden, cf Fig. 27.6)

86
Focus the specimen. It may be easier to focus a Swing in the condenser top (48.15). Engage the
stained specimen first or the edge of the cover- Bertrand lens (50.2) or use the auxiliary
glass. Set Koehler illumination exactly (see telescope (Fig. 51).
page 69), then find an empty area of the Engage the condenser-side prisms (60.8) in
specimen or remove the specimen. succession and focus the dark diagonal com-
Turn the polarizer (60.4) round the IC position pensation stripe. The whole-wave compensator
until the optimum extinction position is observed must be inactive, i.e. the engraving must be on
through the eyepiece. This setting can be found the side of the analyser that points downwards.
particularly accurately with a high-magnification The dark stripe should be in the centre of the
objective (40x or 63x): brighter circular area. If not, proceed as follows:
Open the aperture diaphragm (48.21) as far as push in the right-hand centering key on the back
the stop, engage the Bertrand lens (50.2) and of the condenser until it clicks into position and
focus (50.3) or use the auxiliary telescope (Fig. rotate it until the stripe is in the centre of the
51) instead. The polarizers are exactly crossed circle. The left-hand key is not required.
when the two branches of the hyperbola are as However, for the 3rd and 4th prism position
near to each other as possible – or form a cross make sure that the left-hand centering screw for
(55a). the light rings is not rotated too far inwards or it
Fix this crossed position with clamp screws may obstruct the movement of the prism with
(60.4 and 30.6). the right-hand key.
Put the centering key (1.5) in the index
adjustment (60.5) and make the two index marks Ojectives for ICT
(28.4) coincide; you will now be able to
Transmitted light interference contrast is
reproduce the current polarizer setting later.
possible with the brightfield and phase contrast
objectives which have the code letter of the
Adjustment of the condenser prisms
pupil position in the first line of engraving, e.g. A
If the equipment was delivered together, the and which are listed under the objectives
condenser prisms will already have been suitable for ICT on the optics data sheet.
adjusted at the factory, but it is advisable to Transmitted light interference contrast is also
check the adjustment from time to time, possible with certain incident light objectives
especially after transport: (see separate objective table). A condenser
Disengage the objective side ICT prisms (60.7) prism, e.g. K1 must also be available for the
(pos. H). objective.

87
Choice of prisms (objective prism roughly at the centre position).
Optimum contrast for specimens with parallel
Choose the objective side prism (60.7) with the
structures can be obtained by rotating the stage
letter indicated in the top line of the objective
(48.8).
engraving* (page 48 and on Optics data sheet),
e.g. A for pupil position A.
Colour contrast: Turn over the analyser, so that
Additional numbers e.g. B2: Prism with greater
the sign can be seen on the top. If using the 360°
beamsplitting than the standard version (= B1),
rotatable analyser (30.1) colour contrasting is
for higher detection sensitivity.
carried out by placing a rotatable whole-wave
Choose the condenser side prism (60.8) that
compensator (57.1) on the polarizer or pushing it
corresponds to the magnification of the
into the mount underneath the condenser (27.6)
objective used, e.g. pos. 20/40 for objective 20x
and rotating.
(and 40x).
Swing in condenser top 0.90 S1, only swing out
Specimen preparation
condenser top for the 5x objective (only with
UCR/UCPR condenser; ICT is only possible with ICT gives best results for unstained, relatively
the UCE condenser from objective 10x upwards). thin, non-birefringent specimens. Interpretation
Exactly set Koehler illumination (see page 69). of birefringent specimens can be extremely
This is made easier by temporarily focusing a difficult, if not impossible. It may be helpful to
stained specimen or the edge of the coverglass. rotate the specimen to an optimum azimuth
position.
Setting ICT contrast
Carefully turn the objective-side prism turret
(60.7) to the left and right. Also adjust contrast
with the aperture diaphragm (48.21). Particularly Specimen slides, coverglasses and embedding
sensitive setting is possible with the λ/4 com- resins of birefringent material may not be used.
pensator (57.1), which is inserted in the holder
under the condenser (60.9) and rotated

88
Preparation errors
Errors in instrumentation
Possible sources of error if ICT image is
unsatisfactory: Polarizers not engaged, or rotated too far out of
Embedding medium, specimen slide (Petri dish) the crossed position or, though crossed, turned
or object (e.g. crystals, fibres) are of birefringent out of the zero positions.
material. The phase shifts caused by bire- Polarizer has been damaged by powerful light
fringence disturb the inter-ference contrast sources. Check this by holding the polarizer
image. This can sometimes be remedied by against a window or light source. Damaged
rotating the specimen. polarizers then show distinctly uneven
The specimen is too thick or too thin. colouring.
The IC prisms in the condenser are in the wrong
Specimen slide or coverglass are too thick position or upside down. This is checked by
or the coverglass is missing (except for combining an IC prism with all available
HC PL FLUOTAR 5x/0.12 and 10x/0.25, which can objectives and seeing if the interference
be used either with or without a coverglass). contrast image is optimal at corresponding
magnifications of the objective and the
The difference in the refractive indices of the condenser.
specimen and the embedding medium is too Condenser top in wrong position.
small (this often happens when uncovered Wrong condenser top (only 0.90 S1 or P 0.90 S1
specimens are observed with an immersion or P 1.40 OIL may be used).
objective). Inhomogeneous mounting medium. Koehler illumination not set (image of field
diaphragm in the specimen plane).
Aperture diaphragm too wide or too narrow.
Dirty optics or polarizer.
Dust protection: Turn the condenser prism out of
the light path if it is not being used for a long
time.
For specimens with parallel texture: specimen is
in wrong azimuthal position (remedy: rotate
specimen with stage 60.2; 54.15).

The following description applies for

fluorescence, brightfield, darkfield, polarized
light and interference contrast techniques.

89
 Or:
Back reflection via the specimen
Attention: Focus a well-reflecting incident light specimen
(e.g. surface mirror) (this is not possible for
Never look into the direct light path! There is fluorescence). Remove an eyepiece from the
danger of glare when switching to the tube, or engage Bertrand lens (50.2 or 54.2/11)
brightfield reflector (BF) or the Smith and focus, or use an auxiliary telescope (51.1).
reflector (6.4; 6.5)! The light source can be observed inside the
bright circle (objective pupil) through the tube.
Imaging the light sources to check adjustment
Centration of reflected light lamps
There are several different ways of imaging the
lamp filament or discharge arc; the field Lamphousing 106 (Fig. 4, page 12) with 12 V 100 W
diaphragm (63.6 or 65.8) is first narrowed and the lamp.
aperture diaphragm (65.12) opened. Switch to Adjust the collector (48.19) until you see the lamp
the light source you want to use (61.7*). filament (Fig. 47, page 68).
Using the centering aid
To do this the left side of the microscope stand
must be equipped with the adjustment window
(61.9) for imaging the light source. Put the Using a screwdriver (1.1) adjust the vertical
centering aid (reflector for lamp adjustment, position (48.17) of the lamp holder until the
18.2) into the reflector turret instead of a filter slightly brighter stripe in the reflection of the
cube or reflector (see page 26). Rotate the turret lamp filament is in the centre of the brighter
until the centering aid is in the light path. area (Fig. 47). Then move the reflection of the
Alternatively: lamp filament with the horizontal adjustment
Projection on the microscope base (48.18) to the centre of the range of movement
Remove the specimen and the condenser. Turn (Fig. 47).
in a 5x (or 2.5x) objective. Put a piece of blank
paper or card on the microscope base: the Lamp housing 106 z with halogen lamp and gas
bright circle projected onto it represents the discharge lamp (Fig. 5, p age 13 and Fig. 61)
(unfocused) projection of the objective pupil (in The image of the light source is focused with the
principle the condenser could be left on the collector (61.6) and the holder with the light
microscope and the adjustment carried out source adjusted vertically and horizontally (61.1
through the condenser at a certain setting, but and 61.2). The reflector is also focusable (61.4)
this method is not recommended due to the and centerable in x and y direction (61.3
necessity for exact condenser adjustment). and 61.5).

90
The adjustment principle is similar for all light
sources:
Caution:

Caution with Hg and Xe lamp:

Be careful not to project the reflection on the
Move the reflection of the lamp filament or dis- electrodes for long, as there is a risk of ex-
charge arc to the side or completely out of the plosion if they overheat. The two electrodes
light path (62a) by turning the adjustment can just be seen in the extension of the
screws on the back of the lamphousing (61.3 and symmetry plane of the discharge arc.
61.5). Focus the direct image of the filament or
discharge (61.6) and adjust as follows (61.1, 61.2
and 61.6):

Halogen lamp: just below or above the imaginary Replace spent burners in good time and
line through the centre of the brighter circle dispose of in an environment-friendly way.
(62b). Do not open the lamphousing until the lamp
First focus the reflection (61.4) and then move it has cooled down and you have disconnected
symmetrical to the direct image inside the it from the mains. Wear protective clothing
brighter circle (62c), or superimpose it on top of (gloves and mask) when using Xe lamps. Hg
the direct image. lamps take a few minutes to reach their full
Mercury- (Hg) and xenon lamp (Xe) intensity; they do not ignite when hot.
Move the direct image (62a) to the centre of the
brighter circle with the horizontal (61.2) and
vertical (61.1) adjustment of the holder. Focus the
reflection (61.4) and adjust the mirror until the
reflection coincides with the direct image (62c). Fig. 61 Lamphousing 106 z
1 Vertical adjustment of lamp, 2 Horizontal adjustment of
lamp, 3, 5 Vertical and horizontal adjustment of reflection,
4 Mirror focusing, 6 Collector (focusing of lamp image),
7 Aperture for switch rod* (switchable mirror only),
8 Analyser*, 9 Adjustment window*

91
Collector setting, diffusing screens can be engaged (only with halogen lamp) to
check whether the image is homogeneously
Halogen, Hg and Xe lamps:
illuminated (use a homogeneous specimen
Adjust the collector (61.6) until the bright area is
section if possible and a low-power objective).
uniformly illuminated. Switch the microscope
Use a grooved diffusing screen for the Xe 150 W
back to object observation; the diffusing screen(s)
lamp.

Fig. 62 Lamp adjustment

a direct filament image focused, but decentered
b direct filament image in the right position
c reflected and direct filament image in the right position

a b c

Halogen
lamp

Hg 50
lamp

Hg 100
and Xe
lamp

92
Filter cube, objective, tube factor Set the aperture diaphragm: Remove the
objective and focus a light source on dark paper
Focus the specimen in transmitted light first if
(specimen stage). Narrow and open the dia-
possible. Select a filter cube to suit the
phragm: if the image of the diaphragm lies
excitation and emission spectrum of the
eccentric to the circle: insert the centering keys
specimen and switch it into the light path (63.1),
(23b.8) and center the diaphragm image. Open
see page 26 for assembly. Use high-aperture
the aperture diaphragm in fluorescence mode,
objectives (immersion) to obtain optimum image
only narrowing it in special cases to influence
intensity; open the iris diaphragm in the
contrast. Position of the lever (23.13) for auxiliary
objective if applicable (41.3). Switch the tube
lens (can only be set after pulling the diaphragm
system* to factor 1x. Protect the immersion oil
module HC F out of the microscope):
from impurities to avoid disturbing fluores-
cence. Push in: Optimisation for fov 20 and 22,
gain in intensity (TV!)
Pulled out: Optimization for fov 25
Diaphragm module HC F
Push the diaphragm module in fully (Fig. 63.5 – 10).
Unblock the incident light path (63.8), focus the
specimen and switch off or cover transmitted
light (Fig. 64).
Set the field diaphragm: Close (63.6) until it is
visible in the microscope field of view (49b).
Insert the two centering keys (1.5) in their holes
(63.5) and turn until the image of the field
diaphragm is in the middle of the field of view
(49c). Open the diaphragm until the entire field Fig. 63 Controls for fluorescence with diaphragm module HC F
of view is homogeneously illuminated (49d). 1 Turret for 4 filter systems/reflectors, 2 Interference contrast
If you have an interchangeable stage the cen- prism turret*, 3 Tube lens 1x/Bertrand lens (B)*, 4 Incident
tering keys can be kept on the right side (as in 13.3). light polarizer* mount, 5 Holes for centering keys (field
Disengage the BG 38 filter (63.7) if there is no diaphragm), 6 Field diaphragm, 7 BG 38 filter, 8 Interruption of
the incident light path, 9 Hole for aperture diaphragm
disturbing red background. Always engage the centering keys, 10 Aperture diaphragm, 11 Filter magazine
filter for photography, however. Always dis-
engage the incident light polarizer (63.4).

93
Fluorescence with diaphragm module HC RF: As Possible errors
a BG 38 filter is not integrated here, it must be
Weak fluorescence, weak image intensity due
built into the filter magazine (65.13) if needed.
to:
The light path can be blocked by pulling out the
Incorrectly stored, too old or faded specimens;
diaphragm module part way. Intensity can be
fast specimen fading (e.g. with FITC); inspecific
increased by interposing the illumination
filter combination, numerical aperture of
telescope (“booster”, not illustrated).
objectives too low; eyepiece magnification too
high; spent lamp; room too bright.
Light trap
Low contrast image due to:
To avoid stray light from the underneath of the Excitation bandwidth too great; inspecific
specimen: remove the condenser and put the staining; fluorescing inclusion medium; auto-
light trap (Fig. 64) in its place. Alternatively, a fluorescence of the objective or immersion oil.
black metal plate can be pushed into the stage. With double fluochroming, green and red image
details visible at the same time due to:
Filter cubes unsuitable for selective
observation.
Inhomogeneous illumination due to:
Incorrect lamp centration or flickering lamp.
Brightening of image background or red back-
ground due to:
Absence of BG 38 red attenuation filter in light
path.

Metal staining
The case is different for reflecting objects, such
as in the immunogold technique (IGS). Here the
Fig. 64 Light trap for fluorescence microscopy (instead of the POL filter system (crossed polarizers for
condenser). Instead of the light trap, a dark metal or plastic contrast enhancement) is used for incident light
strip can be used, which is inserted between the upper and
lower part of the x/y stage under the specimen.
polarization instead of a fluorescence filter
cube, and contrast, resolving power and depth
of field can be influenced with the aperture
diaphragm.

94
Reflection contrast* Polished specimens can be pressed plane-par-
allel onto a metal specimen slide (code no.
The following equipment is required (see sepa-
563 014) with the special handpress (code no.
rate manual):
563 035) and plasticine. The handpress has an
Reflector system POL (Fig. 18), special objective
adjustable stop so that all specimens can be
RC with rotatable λ and λ/4 compensator
aligned to the same height. Then only slight
mounted in front, reflection contrast module
refocusing is required with the fine control
HC RC, with additional annular diaphragms for
during serial investigations.
optimising contrast.
Objects that do not lie flat and that cannot be
levelled with the handpress can be aligned by
Incident light brightfield*, alignment of
autocollimation. The object is focused on the
polished sections
tiltable specimen slide, for example, (code no.
Homogeneous illumination and uniform 562 294) at a low magnification (5x or 10x
definition over the whole field of view can only objective).
be guaranteed if the surface of the specimen is
aligned at exactly 90° to the optical axis. A
precisely horizontal position of the object is
particularly important at high magnifications, as
the depth of field decreases as magnification
increases.
Fig. 65 Controls for incident light brightfield, darkfield,
polarized light, ICR interference contrast, see also Fig. 23
1 Analyser (hidden, on the left of the microscope, cf Fig. 30),
2 Tube lens 1x/Bertrand lens*, 3 4-position turret* for
reflectors/filter system, 4 Incident light polarizer*, 5 Turret for
objective-side Wollaston prisms* or Pol compensator slit,
6 Contrast adjustment ICT and ICR, 7 Holes for centering keys
(field diaphragm), 8 Field diaphragm, 9 Switch lever on
diaphragm module HC RF, 10 Grey filter, 11 Aperture
diaphragm (hidden), 12 Aperture diaphragm decentration
(oblique light illumination), 13 Centration of aperture
diaphragm, 14 Filter magazine*

95
The field diaphragm must also be exactly Diffusing screen pairs A and B
centred in the field of view (65.7) and the
The diaphragm module HC RF is equipped with
aperture diaphragm (65.11/12) closed.
an interchangeable pair of diffusing screens
Remove the objective or objective nosepiece:
(23b.9) to obtain optimally geneous illumination
the reflection of the field diaphragm is reflected
both for visual observation and for video and
in the centre of the field of view if the object
digital image processing.
surface is aligned exactly horizontal.
Diffusing screen pair A is included in the
standard delivery and contains diffusing screen
Diaphragm module HC RF
1 with dense distribution for even mination over
With 2 illumination channels and interchangeable a large field of view of 25 mm or 28 mm with the
diffusing screens for incident light BF, DF, POL, DM RD HC.
ICR, FLUO. Diffusing screen 2 with low distribution for
maximum illumination homogeneity, but over
Illumination channel I a reduced field of view of max. 20 mm (video
(Diaphragm module fully pushed in) and digital image processing). Diffusing screens
For use in all incident light modes with 1 and 2 can be used in either of the two
– variable iris aperture and field diaphragm illumination channels. To do this, remove the
– oblique illumination diffusing screen pair, which are held by
– switchable neutral density filter magnetism, and turn by 180° so that the labelling
A, 1 2 is upside down.
Illumination channel II
(Diaphragm module pulled out as far as the 1st
clickstop)
For use in all incident light modes with
– fixed aperture and field diaphragm
– mount for focusing graticule

Illumination channel II offers the additional

advantage of fast, reproducible switching
between open and closed diaphragms e.g.:
If the diaphragm setting is to be retained when
switching between brightfield and darkfield.
For fast switching between high and low
objective magnifications.
Channel II is also advantageous for measure-
ments with fixed diaphragms, for colour assess-
ment of coatings and oxide films and for work
with the focusing graticule.

96
As well as diffusing screen pair A, 1 2 diffusing Open the field diaphragm (49d) until it just
screen pair B, order no. 565 502, can be disappears from the field of view.
supplied. Diffusing screen pair B contains 2 This setting of the field diaphragm is retained for
identical diffusing screens and is recommended all objectives.
when the same illumination conditions are If the diaphragm module HC RF is pulled out
required on both channels. as far as the st clickstop (= channel II), the field
and aperture diaphragms are fixed, see chart on
Incident light brightfield p. 96.
Set the microscope illumination to medium
The field diaphragm only has to be readjusted
intensity (42.14 and 42.8).
when eyepieces with different field numbers are
Turn in a low power objective (e.g. 10x). Make
used, when the secondary magnification is
sure the front lenses of the objectives are clean!
altered with a magnification changer or zoom
Push in the diaphragm module (65.9) as far as
system, or for photography and filming.
the stop (= channel I).
Narrowing the field diaphragm usually improves
Close the field diaphragm (65.8). Open the
the contrast.
aperture diaphragm (65.12).
For interchangeable stages only: The centering
Using the stage clamp (48.9) and the coarse
keys can be kept in the stage bracket (42.11 or
focus control (42.12) or (44.2 and 44.3), position
13.3) after use.
the sample surface roughly in the focal plane (=
45 mm below the objective thread, see page 57).
The aperture diaphragm (65.12) affects
Focus the object. The image of the closed field
resolution, contrast and depth of field of the
diaphragm (65.8) makes it easier to find the
microscope image.
object surface.
It must be set carefully and must not be used to
See page 67 for tube and eyepiece setting.
adjust image intensity.
Setting the field diaphragm:
Close the field diaphragm (65.8) until its edge
can just be seen within the observed object field
(49b). Put the two centering keys (1.5) into the
holes (65.7) and adjust until the edge of the field
diaphragm is concentric with the edge of the
field of view (49c). Centration of the closed field
diaphragm can also be performed with a
graticule e.g. with crosslines.

97
Engage the Bertrand lens (50.2) and focus (50.3) Incident light darkfield
or remove an eyepiece and look into the tube
Special darkfield objectives (BD, Fig. 40) with
from a distance of a few centimetres. Mount the
built-in annular mirror or annular lenses are
centering keys (23.8) and adjust so that the
required for incident light darkfield. These
closed diaphragm lies in the centre of the
objectives have a greater external diameter and
brighter circle (= objective pupil). Open the
screw thread M32 x 0.75.
aperture diaphragm until it is just visible in the
High light intensity is necessary for darkfield, as
brighter circle (= objective pupil). The
this type of illumination is produced by diffracted
illumination aperture is then equal to the
and scattered light. Therefore, remove all filters,
observation aperture.
polarizers, IC prisms, etc. from the light path and
After returning to the normal observation mode
set maximum intensity.
(Bertrand lens disengaged) image contrast can
be individually adjusted.
If the aperture diaphragm is stopped down too
far – especially at low and medium objective
magnifications – the object structures will Make sure the front lens of the objective is
exhibit pronounced diffraction phenomena. clean as this has a great influence on the
The aperture diaphragm can be stopped down imaging quality in darkfield.
further for high-power objectives to improve
contrast and depth of field. Fine-adjust the Pull out the incident light diaphragm module
aperture diaphragm, watching the structure and HC RF (65.9) as far as the first (= channel 2) stop.
topography of the object, to obtain the best The aperture and field diaphragm functions are
contrast and resolution. then set.

A neutral density filter (65.10) can be engaged in

the light path to adjust the image intensity when
switching to brightfield.
This neutral density filter is only in channel I. It
saves the user from having to reduce the lamp
intensity and is particularly useful when
switching quickly between techniques DF ↔ HF.

Oblique light
For brightfield illumination the illuminating cone
is rotation-symmetrical to the optical axis. For
oblique light the aperture diaphragm (65.11 and
65.12, for channel I only) is moved to the side and
stopp ed down so that the illuminating cone hits
the sample at an angle, highlighting the surface
topography.

98
Incident light interference contrast ICR Contrast setting
Carefully move the turret round the centre
Cross polarizers.
position, additionally operating the aperture
Exactly crossed polarizers are an essential
diaphragm (65.12) to optimize contrast.
requirement for perfect ICR quality!
Insert the ICR polarizer (29.5) (65.4). Never use a
The interference contrast technique gives a
different incident light polarizer. Switch on
relief-like and three-dimensional image of the
brightfield reflector BF or Smith reflector (Fig.
specimen surface.
18); push in diaphragm module (65.9, channel I).
The contrast of linear structures can be
Turn the turret* (65.3) to pos. H (= brightfield).
improved even more by rotating the specimen
Align and focus a homogeneous and well-
with the stage rotation control (48.8).
reflecting specimen.
For observations in ICR colour contrast, remove
Insert the IC/P analyser so that the lambda (λ)
the analyser slide, rotate by 180° and push back
engraving is showing (65.1 and 30.5). If using the
in with the λ engraving showing. In this position
360° rotatable analyser (30.1) set the 0 position.
a whole-wave compensator is effective in front
Swivel the analyser round the 0 position until
of the analyser, producing colour interference
you obtain the darkest possible setting.
contrast. With the analyser 360 and the ICR
Instead of the polarizer and analyser, the ICR
module, colour contrast is only possible with the
module (= crossed polarizers) can be used
“turnaround” polarizer L ICR with whole-wave
compensator.
Choice of IC prism
To switch from interference contrast to bright-
Choose the prism in the turret (65.5) that
or darkfield, turn the prism turret to position
corresponds to the objective you are using, see
H = brightfield, and pull out the polarizer and
objective engraving, p. 48, or “Optics” data
analyser by one clickstop.
sheet. Optionally an IC prism in slide mount can
be inserted in the Pol compensator slot (54.13).

Prisms with an additional number, e.g. D1, B2,

split the beam than the D or B1 prism and are
therefore more sensitive for detecting fine
topological details. However, the B1 prism is
used for obtaining the highest possible
resolution.

99
Quantitative interference attachments
Surface roughness and topography are depicted Setting the R/P polarizer (29.1):

↔
as interference fringes by the various When combined with IC/P analyser (30.7)
interference techniques. These are evaluated ↔ When combined with 360 analyser (30.1)

similar to the way contour lines are interpreted

on maps. The measurement accuracy is up to Focus an isotropic specimen that fills out the
30 nm, the maximum height difference is about whole field of view, e.g. a mirror, open the
30 µm. See special instructions. aperture diaphragm (65.12) and turn the analyser
(65.1) until the maximum extinction position can
Fibre-optic, light guides be seen. For the IC/P analyser (30.5) the λ
engraving must point downwards (compensator
Illumination with flexible fibre-optic light guides
inactive).
with ball-jointed arms (VOLPI intralux 6000),
As for transmitted light (page 77) a particularly
rotatable round the optical axis of the
precisely crossed position can be achieved with
objectives. Colour glass filter(s), slip-on iris
a Bertrand lens or auxiliary telescope.
diaphragms, auxiliary lenses, polarization
device (polarizer and analyser).
Polarizer with rotatable whole-wave compen-
sator (29.9)
Set the analyser exactly at 0° or 90°.
Incident light polarization
Turn the whole-wave compensator (29.3) roughly
Adjustment to the centre position.
Set the light source and diaphragms as for Turn the polarizer until the object appears as
incident light brightfield (page 90 and 95). dark or as highly contrasted as possible, turn the
Reflector: BF or Smith, the Smith reflector is whole-wave compensator (29.4) until colour
better from a polarization optic point of view and contrast is obtained.
should be used for slight anisotropy (polari-
zation) (see Fig. 30). Filter system POL (p. 33 and ICR)
This does not need adjusting, as polarizer,
Crossing the polarizers analyser and 45° flat glass reflector are
Important: The polarizers should be exactly combined as a fixed unit.
vertical or horizontal as a deviation of even 1°
may lead to impaired extinction.

100
Possible errors, brightfield, darkfield, ICR Inhomogeneous image illumination:
Lamp not adjusted.
Fall-off of focus, one-sided:
Sample not aligned flat.
Sample surface not aligned at exactly 90° to the
optical axis.
For bright-/darkfield:
Sample has round edges.
Oblique illumination lever not in exact position.
Stage not clamped tight.
Diaphragm module not in exact position.
Polarizer/analyser slide not exactly positioned.
Fall-off of focus on both sides:
Tube beamsplitter at incorrect setting.
Sample surface greatly inclined.
For ICR interference contrast:
Fall-off of focus, partial:
IC prism in light path.
Pronounced relief zones in the sample beyond
Wrong IC prism engaged.
the range of the depth of focus of the objective.
Polarizer discoloured due to overheating.
Incorrect polarizer setting (see page 100).
Fall-off of focus, concentric:
Sample surface is round.
Object marker
Image is unusually flat: The object marker is screwed in place of an
Poor sample quality. objective (not illustrated). When rotated, a
Fingerprints or dirt on front lens of objective. diamond is lowered onto the coverglass or
Sample covered by other layers. object surface, where circles of variable radii
Illumination aperture not exactly matched to the can be scribed to mark objects.
sample (Close aperture). Multi-viewing attachment: see separate manual.
Objective in use is not suitable for reflected light
(see DELTA optics objective data sheet).

101
Diapositive overlay To do this, the original must be photographically
reproduced on a 35 mm negative, i.e. bright lines
The diapositive overlay device (Fig. 66) is used
on a dark background and framed in a standard
to reflect measurement and comparison masks,
50 x 50 mm slide frame. The best film to use is
µm marks, marker arrow, company logo, charge
fine-grain “document film”.
and quality data, etc. into the microscope image
The diapositive is imaged in the intermediate
so that they can be recorded together with the
image plane of the microscope at a scale of 2 : 1.
image. Only with tube HC FSA 25 PE and with
A distance of e.g. 5 mm in the diapositive overlay
DM RD (Fig. 31 and 33)!
is magnified to 10 mm in the intermediate image
The following diapositives are available:
plane of the microscope. The overlay device
only works when the beamsplitter in the tube
Marker arrow
(31.4) is set at 50/50 (switch rod in middle
10 mm measurement scale with 100 divisions
µm marks for 2.5x – 100x objectives position). The framed diapositive is inserted in
10 x 10 mm grid division in 100 fields the integrated holder (66.6) (white side of
Test circle and measured length for grain size diapositive with lettering facing microscope).
measurements
Picture series for ASTM-E 112 grain size
measurements.

Individual masks with any measurement and

comparison patterns, quality data, company
logos etc. can be made by the user.

Fig. 66a Diapositive overlay device on the HC FSA 25 PE tube

1 Tube flange, 2 Coupling ring for reflection optics, 3 Reflec-
tion optics, 4 Coupling ring for diapositive overlay device,
5 Knurled ring for focusing, 6 5 x 5 cm slide frame, 7 Filter slot,
8 Illumination tube of lamphousings Fig. 66b Transformer

102
The holder is adjustable on all sides, so the The image is observed in the microscope tube
overlay can be moved to different areas of the and focused by turning the knurled ring.
microscope image. Remember that when you The magnification can be changed continuously
move the diapositive, the overlay will move in in a range of 1 : 4 by adjusting the knurled ring
the opposite direction. This takes a bit of getting (67.7).
used to. When changing the magnification with the zoom
You can give the bright lines a coloured back- control the object may change position slightly
ground by putting 32 mm colour filters in the and go out of focus. It must then be refocused
filter slot (66.7). and moved back into position.
The zoom magnification factors can be read on
Macro device the scale (67.8). The magnification also changes
when the distance between the object and the
Like the diapositive overlay device, the macro
macro attachment is varied.
overlay (Fig. 67) only works in the 50/50
beamsplitter position (switch rod at middle
position) of the HC FSA 25 PE tube.
The microscope illumination is left switched off
to avoid disturbing image brightening.
The object is placed on the stage under the
mirror housing of the macrodual zoom (67.11) and
illuminated.
Stand lamps, cold-light illuminators and fibre-
optic lamps, etc. are suitable light sources for
macroscopy.

Fig. 67 Macro device on HC FSA 25 PE tube

1 Tube flange, 2 Coupling ring, 3 Reflection optics, 4 Coupling
ring, 5 Macro adapter, 6 Screw ring, 7 Zoom setting ring 1 : 4,
8 Scale of zoom factor, 9 Scale of magnification factor of the
working distance, 10 Scale of object distance from the bottom
edge of the mirror housing, 11 Mirror housing

103
The total magnification in the microscope, the The total magnification in the film plane of a
reproduction ratio on the photograph or TV camera is derived from multiplying the
image can be quickly and easily measured with intermediate image magnification M1 by the
a scale and calculated. magnifications of the photo eyepiece and
camera attachment, e.g.:
Important: For normal viewing without the macro
mirror housing or macrodual zoom, put on the intermediate image magnification 0.125x
cover to avoid disturbing overlay effects. photo projection lens 8x
large-format attachment 1.25x
The mirror housing (67.11) can be rotated through 0.125 x 8 x1.25 = 1.25x
360°, for example to alter the angle at which the The total magnification at the 4 x 5′′ large-format
photograph is taken. This is done by loosening camera of the DM LD would therefore be 1.25x.
the Allen screw.

The intermediate image magnification M1 the

macro object can be worked out from the
eyepiece field of view (see page 43) and the
diameter of the object field (measured with a
graduated ruler) as follows:
M1 = z. B.
field of view Ø e.g. ––––––––––––––––––
M1 = ––––––––––––– 10x/25 eyepiece M = 0.125
object field Ø object field = 200 mm
M1 = M1 =

Viewed with a 10x eyepiece, this intermediate

image of 0.125x gives a total magnification of 1.25x
in the microscope eyepiece (0.125 x 10 = 1.25).

104
The total magnification can be roughly worked The exact magnification of the object in the
out using the scale divisions on the macrodual drawing is most easily determined by means of
zoom: a stage micrometer, by transferring the length
measured by the stage micrometer onto the
The following factors are multiplied: drawing. The magnification can also be
– magnification factor of the working distance calculated as follows:
(scale 67.9, e.g. – 0.11x) MZe =
MObj 5x
– zoom factor (scale 67.8, e.g. 1x) MZe = ––––––––––––– e.g. ––––––––––––– = MZe 9.6x
FZoom x FD x FE 4 x 0.11 x 1.176
– correction factor of the reflection optics MZe =
(without engraving 1.17x) MZe = magnification in the drawing plane
– eyepiece magnification (e.g. 10x) MObj = objective magnification
e.g. 0.11 x 1 x 1.17 x 10 = 1.29 FZoom = magnification factor of the zoom optics,
The total magnification in the eyepiece would scale 67.8.
therefore be 1.29x. FD = magnification factor of the object
distance, scale 67.10
Using the macrodual zoom as a drawing device FE = correction factor of the reflection optics
Drawing microstructures under the microscope (1.176x)
has the advantages over photomicrography that
significant details can be high-lighted and that The magnification can be altered by changing
structures can be depicted in three dimensions. the zoom setting (scale 67.8) or the level of the
Apart from this, drawing with the superimposed drawing plane.
image method is a valuable didactic exercise.
It is done by superimposing the drawing area
(the area of the stage under the mirror housing
of the macrodual zoom) onto the microscope
image. The drawing area or sheet of paper is
homogeneously illuminated with a stand lamp or
table lamp.
The microscope illumination and illumination of
the drawing area are matched providing the
lamps are adjustable; otherwise the brightness
of the drawing area can be varied by altering the
proximity of the lamp.

105
At the smallest zoom setting the drawing area Linear measurements
has a diameter of approx. 190 mm, at the highest
The following are required for linear measure-
zoom setting approx. 48 mm with an eyepiece
ments:
field of view of 25 mm. For different fov numbers
– Graticule with scale division in eyepiece
the correction value is fov/25.
(Fig. 68) or Variotube DM RD HC (Fig. 33) or
diapositive overlay device (Fig. 66) or a digital
Auxiliary lens 2x
linear measuring eyepiece.
An auxiliary lens 2x can be screwed in under the – Transmitted or incident light stage micrometer
mirror (67.11) to magnify the field that is to be for calibration.
imaged. This must be taken into account for the
above formula. This auxiliary lens 2x is The micrometer value of the objective-eyepiece
recommended for microscopic tracing as object combination used must be known before the
structures are shown twice as large. measurement, i.e. the distance in the specimen
that corresponds to the length of a division on
Overlay of data and code numbers with the the graticule used.
VARICODE systems
The VARICODE system can be supplied together
with the macrodual zoom.
It allows code numbers, micron measurement
bars, ASTM grain size pictures and 35 mm nega-
tives to be overlaid on the microscope image.
Further details on how to use this system can be
found in the manual of the manufacturer, Leica
AG, Vienna. Not illustrated.

VARIMET digital measurement system

The VARIMET measurement system can be
connected to the reflection optics for the
measurement of microstructures. An adapter is
available on request. See manufacturer’s
manual (Leica AG, Vienna) for further details.

106
Calibration: Important: If using a Variotube or variable tube
Align the stage micrometer and the graticule factor:
parallel to each other by rotating the stage or Remember to take the additional magnification
the eyepiece and adjust the zero marks of both value into consideration! We strongly
scales to exactly the same height. recommend you calibrate each objective
separately instead of extrapolating the
Read how many scale divisions of the stage micrometer values of the other objectives from
micrometer correspond to how many on the the calibration of one objective. Measurement
microscope scale (graticule) and divide the two errors may occur if the eyepiece is not pushed
values. into the tube as far as the stop.
Particularly large object structures can also be
Example: measured on the stage with the verniers
If 1.220 mm of the stage micrometer corresponds (0.1 mm); the distance to be measured could
to 50 divisions of the measurement scale, the be calculated from a combined x and y
micrometer value is 1.220 : 50 = 0.0244 mm = 24.4 measurement.
µm. For extremely low objective magnifications
it may be that only part of the measurement
scale can be used for calibration.

Fig. 68 Graticule division in eyepiece (left) and image of the

stage micrometer (right)

107
Microscopic measurement and comparison in The graticule for ASTM-E 112 grain size
metallography measurements is divided into eight segments
Linear measurements with measurement grati- with numbered grain sizes. The pictures
cules conform with grain size plate no. 1 of the ASTM-
E 112 standard. We refer to the ISO/DIS 643,
The size of the line patterns and the length of the Euronorm 103/71, DIN 50 601 and ASTM-E 112
divisions are designed for the standard standard specifications for taking grain size
magnifications customary in metallography. For measurements with the named graticules.
standard magnifications one graticule division Digital length and height measurement using TV
has the following rounded values in the object technology: see separate Leica MFK 2 manual.
plane:
standard magnification: Thickness measurements
100x – 1 division approx. 10 µm
In principle, thickness measurements can be
standard magnification:
carried out if both the upper and the lower
200x – 1 division approx. 15 µm
surface of the object can be clearly focused.
standard magnification: 1
The difference in stage height setting
500x – 1 division approx. 12 µm
(mechanical dual knob focusing: distance
standard magnification:
between two divisions = 2 µm) gives a value for
1000x – 1 division approx. 11 µm
transmitted light objects that is falsified by the
refractive index of the object (which has been
The exact proportions of measurement divisions
“transfocused”) and perhaps immersion oil. The
in the microscope can be checked by using
true thickness of the object detail measured in
stage micrometers, calibration standards or
transmitted light is given by the vertical stage
microscales.
movement (focusing difference) d’ and the
refractive indices no of the object and ni of the
Graticules for grain and particle size determination
medium between the coverglass and the
The graticules for the standard series and
objective
Snyder Graff methods contain a test circle
d = d’
which the viewer sees as having a diameter of no
d = d’–––
ni
80 mm at standard magnification. Its size
d = d’
therefore conforms with the standard picture
series charts, facilitating size comparison.
These graticules also include a measured length
to allow the Snyder Graff line sectioning
method. This and similar methods involve
counting the number of grains cut by the
measured distance. An average grain size can
be worked out by taking several measurements.

108
Example Recorded picture diagonal with
The upper and lower surfaces of a thin polished 1 inch 2/3 inch 1/2 inch 1
/3 inch
camera camera camera camera
specimen have been focused with a dry
objective (ni = 1.0) scale readings of the mechanical Without zoom magnification for 1 chip cameras
c-mount adapter 1x HC 16 11 8 6
fine drive (division spacing = 2 m) : 19.0 and 12.5. c-mount adapter 0.63x HC – 17.5 12.7 9.5
Therefore d’ = 2 x 6.5 µm. c-mount adapter 0.5x HC – – 16 12
The refractive index of the object detail was c-mount adapter 0.35x HC – – – 17.1
c-mount adapter 4x HC 4 2.8 2 1.5
taken to be no = 1.5.
Thickness d = 2 x 6.5 x 1.5 = 9.5 µm Without variable magnification, for 1 – 3 chip cameras
in conjunction with TV optik 0.5x HC (screwed connection)
c-mount adapter 1x – – 16 12
TV microscopy B-mount adapter 1x – – 16 12
B-mount adapter 1.25x – 17.5 – –
Various adapters are available for the F-mount adapter 1x – – 16 12
connection of TV cameras with c-mount or B- F-mount adapter 1x – 17.5 – –
mount objective thread (Fig. 69).
With zoom magnification (Vario TV adapter) for 1 chip cameras
c-mount, 0.32 – 1.6x HC – – 19+) – 5 18 – 3.8
C-mount adapters listed in the following table B-mount, 0.5 – 2.4x HC (SONY) – – 16 – 3.3 –
can be used on all phototubes and on the Leica B-mount, 0.5 – 2.4x HC (SONY) – – – 12 – 2.5

DM RD photomicroscope. The picture area on +)

from zoom factor 0.42x!
the monitor depends on the adapter used and on
the chip size of the camera (s. table).

Fig. 69 c-mount adapter and B-mount (Vario)

1 TV camera, 2 Adapter with c-mount thread, 3 Clamp screw in tube head
a c-mount adapter for 1 chip cameras b Vario TV adapter

109
TV cameras with bayonet mount Incorrect colour rendering
Cameras with the standard Sony bayonet mount Remedy: Vary illumination intensity, carry out
can also be connected to all phototubes, the white balance for TV camera according to
Leica DM RD photomicroscope and the Vario- manufacturer’s instructions, use a conversion
tube 28 VPE. A/B-mount adapter 0.55x and a filter, e.g. CB 12.
Vario B/C-mount adapter 0.55x – 1.1x are
available for this purpose. The recorded field Disturbed picture frame
sizes can be seen in the table. Remedy: Earth the microscope, Variotube and
camera. Avoid parallel laying of mains and
Calculation of the magnification on the monitor signal cables; connect camera and microscope
For all FSA tubes the magnification on the to the same mains phase (socket).
monitor can be calculated with the following
formula: Picture spoilt by inhomogeneous glare and/or
spots. Lamps or windows are reflected in through
VTV = objective magnification x tube factor x TV- the eyep ieces.
Remedy: Switch over the beamsplitter or cover
monitor diameter
adapter magnification x ––––––––––––––––––––– eyepieces or remove the disturbing light source.
chip diameter of camera Dirt particles in the light path, lamphousing not
centered (TV systems are generally more sensi-
If using the magnification changer or the Leica tive to inhomogeneous illumination).
DM RD HC photomicroscope the above formula
must also be multiplied by the factor of the
magnification changer or zoom.

Possible errors
Picture too dim (noisy TV picture, poor contrast)
Remedy: Increase lamp intensity, swing filter out
of light path, switch over beamsplitter in tube
system, switch TV camera to higher sensitivity.

Picture too bright (TV picture glare)

Remedy: Switch neutral density filter, switch
over beamsplitter in tube system, reduce
camera sensitivity.

Picture area too small

Remedy: Use a TV adapter with a smaller factor.

110
Care and maintenance
suction. Any remaining dirt can be removed with
a clean cloth moistened with distilled water.
Attention:
Failing this, use pure alcohol, chloroform or
benzine.
Disconnect from the mains before cleaning
and servicing! Oil
First wipe off immersion oil with a clean cotton
Dust protection
cloth, then wipe over several times with ethyl
Protect the microscope and peripherals from alcohol.
dust by putting on the flexible dust cover after Attention: Fibre and dust residue can cause
each work session. Dust and loose particles of disturbing background fluorescence in
dirt can be removed with a soft brush or lint-free fluorescence microscopy.
cotton cloth. Objectives must not be opened for cleaning.
Only the front lens can be cleaned in the ways
Solvents described above and the upper lens by blowing
Obstinate dirt can be removed with a clean dust off with a bellows ball.
cotton cloth moistened with any ordinary
hydrous solution, benzine or alcohol. Do not use All Leica instruments are manufactured and
acetone, xylol or nitro dilutions. Cleaning agents tested with extreme care. If you do have cause
of unknown composition should be tested on an for complaint, however, please do not try to
inconspicuous part of the microscope. Painted repair the instruments and their accessories
or plastic surfaces must not be tarnished or yourself. Contact your national agency or our
etched. central servicing department, the Technical Ser-
vice in Wetzlar direct.
Acids, alkaline solutions
Postal address:
Particular care should be taken when working Leica Microsystems Wetzlar GmbH
with acids or other aggressive chemicals. Abt. Technischer Service Tel. +49 (0) 64 41-29 28 49
Always avoid direct contact between such Postfach 20 40 Fax +49 (0) 64 41-29 22 66
chemicals and the optics or stands. Thorough D-35530 Wetzlar Telex 4 83 849 leiz d
cleaning after use is strongly recommended.
Keep the microscope optics absolutely clean. Please direct any questions on application to
our Produktmanagement Mikroskopie.
Dust/optics
Remove any dust from glass surfaces with a
fine, dry, grease-free artists’ hair brush, or by
blowing with a bellows ball or by vacuum

111
Main wearing and spare parts, tools
Code no.
Part no. Component Used for
Spare lamps
500 974 halogen lamp 12 V 100 W Lamphousing 107/106 z
500 296 halogen lamp 6V 4W ORTHOMAT E, Leica DM RD,
Diapositive overlay
500 137 max. pressure Hg lamp 50 W Lamphousing 106 z
500 138 max. pressure Hg lamp 100 W Lamphousing 106 z
500 321 max. pressure Hg lamp 103 W/2 Lamphousing 106 z
500 139 max. pressure Xe lamp 75 W Lamphousing 106 z
500 186 Na spektral lamp Lamphousing 106 z
Tools/adjustment key
553 407 Centering keys, short 1.5 x 23 mm Diaphagm module F/RF
020-422.573-053 POL centering keys, Pol microscope
long version (1.5 x 51 mm)
553 143 Centering keys, square Sénarmont compensator
016-500.020-006 3 mm Allen key
150-000.100-129 2 mm Allen key, angled Assembly for light rings and
ITC condensor Wollaston prisms
150-000.100-128 1.5 mm Allen key, angled Torque setting of
x/y-stage movement
Screw cover for unoccupied nosepiece positions
020-422.570-000(4) Screw cover M25 Septuple objective nosepiece
and sextuple centrable
objective nosepiece
016-016.005-200 Screw cover M32 BD objective nosepiece (sextuple)
Spare eyecups (anti-glare protection) for HC PLAN eyepiece
021-500.017-005 Eyecup HC PLAN 10x/25 eyepiece
021-264.520-018 Eyecup HC PLAN 10x/22 eyepiece
021-264.520-018 Eyecup HC PLAN 10x/20 eyepiece

Immersion oil, DIN/ISO standard, fluorescence-free

513 787 10 ml OIL and IMM objectives
513 522 100 ml and oil condenser tops
513 788 500 ml
Spare fuses, primary
846-205.000-000 IEC 127-2 T 4 A all microscopes
832-493.000-000 IEC 127-2 T 2.5 A Xe 75 Hg 100 power unit,
for 90 – 140 V stabilized (500 311)
827-902.000-000 IEC 127-2 T 1.25 A Xe 75 Hg 100 power unit,
for 90 – 140 V/187 – 264 V stabilized (500 311)
824-716.000-000 IEC 127-2 T 0.16 A Xe 75 Hg 100 power unit,
for 90 – 140 V stabilized (500 311)
826-095.000-000 IEC 127-2 T 0.08 A Xe 75 Hg 100 power unit,
for 187 – 264 V stabilized (500 311)
825-347.000-000 IEC 127-2 T 2 A 100 power unit,
non-stabilized (500 299)
The following external power units are without fuses: Hg 50 (500 277)
Xe 150 (500 298)
Hg 200 (500 235)

112
Index
Achromat 48 Filters 16, 17, 56 Parfocality 62
Addresses 2, 111 Flash 11, 16 Phase contrast 23, 50, 73
Adjustment of incident Fluorescence 26 Photo 38
light 27, 57, 90 FLUOTAR® 49 Photo eyepieces 44
Adjustment of Focusing 54, 57 Photomicro 37, 39, 109
transmitted light 57, 65, 87 Focusing graticule 64 Planachromat 48
Adjustment specimen 54 Fuses 9, 10 Planapochromat 48
Analyser 34, 77, 86 Polarized light 77, 83, 100
Aperture 49 Graticules 44, 106 Polarizer 32, 77, 86, 99
Aperture diaphragm 49, 71, 97 Power supply 9
Heating stages 49
Apochromat 49 Power unit 9
Height adjustment of
Auxiliary lens 106 Pupil 48
Condenser 69, 70
Auxiliary telescope 73 Push-on cap 51
Height adjustment of stage 61
Basic setting 53 Hg lamps 13, 91, 112
Quarter-wave compensator 25, 80, 101
BD 50 Quartz wedge 80
Bertrand lens 64, 73, 77 ICR 30
Brightfield 69, 96, 97 ICT 25, 30, 86
IGS 94 Reflection contrast RC 94
Burners 13, 90 Reflections 52
Immersion 50, 65, 73, 76
Calibration 58, 103, 106 Incident light darkfield 98 Reflector 26, 54
Care 111 Incident light illuminator 26, 90 Resolution 49
Centration 65, 68, 90 Installation 8
Circular polarization 80 Interference attachment 52, 99 Safety information 5, 9
Cleaning 111 Interference contrast 25, 32, 86, 99 Service 112
Collector 11, 15, 72, 92 Intermediate rings 45 Spare parts 112
Colour coding 52 Iris diaphragm 49 Specimen 55, 88
Colour temperature 53, 61 Specimen stage 18, 54, 57
Compensators 25, 80 Koehler illumination 69 Stage height stop 61
Condenser 21, 69, 75 Survey observation 21, 64, 80, 102
Lamp adjustment 68, 92 Switching on 53, 58, 61
Condenser height stop 70 Lamp change 12, 41
Condenser holder 19, 20 Length measurement 106
Condenser prisms 25, 87 Technical data 5, 112
Lens (2x) 106 Thickness measurement 108
Condenser top 22, 71 Lens attachments 51
Conformity 113 Tools 8
Light rings 23, 73 Torque adjustment 54
Conoscopy 83 Light sources 10, 68
Contrast 49, 71, 88, 97 Transit protection 19
Light trap 94 Transmitted light darkfield 75
Correction objective 51 Locking of objectives 51
Coverglass 47, 65 Tube 37, 67
Cross section diagram 6 Macro device 41, 103 Tube length 47
Magnification 42, 49, 61, 104 Tube lens 47
Data 5, 112 Tube optics 6, 35, 53, 73, 83
Mains frequency 5, 9
Depth of field 49 TV 109
Mains voltage 5, 9
Diaphragm modules 29, 93
Mercury lamps 13, 91, 112
Diaphragms 49, 69, 83, 93 Useful magnification 43
Mirror housing 10
Diapositive overlay 40, 102
Diffusing screen 72, 92, 98 Object field 43 VARICODE 106
Drawing device 105 Object guide 20, 55 VARIMET 106
Engraving 47 Object marker 102
Objective nosepiece 31, 45, 60 Water immersion 5
Errors 11, 72, 74, 76, 85, 89, 94, 109 Whole-wave compensator 25, 80, 101
Eyepieces 42, 67 Objective prisms 30, 48, 87, 99
Objectives 47, 65, 87 Wollaston prism 88
Field diaphragm 69, 96 Oblique illumination 99
Field of view index 43 Oil 50, 111 Xenon lamp 13, 91, 112
Filter systems 26, 34, 54, 93 Order code nos. 52, 112

113
EU-Confirmaty declaration
EU-Confirmity declaration
We hereby declare that the product specified
below conforms in its design and construction
as well as the model we have put on the market
to the relevant safety and health regulations laid
down by the European Union. This declaration
will cease to be valid if the instrument is
modified without our consent.

Product names: DM R, DM RE, DM RX, DM RXE,

DM RXP
Instrument type: Light microscope
Instrument no.: 020-525.701 to 020-525.780
EU directives: Low voltage: 73/23/EWG
Electromagnetic
compatibility:
89/336/EWG
Harmonised EN 50 081-1
standards EN 50 082-1
applied: EN 61 010-1

Wetzlar, den 18. 4. 1998

Prof. Dr.-Ing. habil. M. Jacksch,

Director of Technology and Development
planning

114