TROUBLESHOOTING THE CURVES

Please select an instrument

CAPILLARYS 2
CAPILLARYS 2 FLEX PIERCING
MINICAP
MINICAP FLEX PIERCING

MEMO
Troubleshooting Curves - 2016 10 17
TROUBLESHOOTING THE CURVES
CAPILLARYS 2
Please select a technique

PROTEIN(E) 6 TECHNIC
HEMOGLOBIN(E) TECHNIC

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TROUBLESHOOTING THE CURVES
CAPILLARYS 2 FLEX PIERCING
Please select a technique

PROTEIN(E) 6 TECHNIC
HEMOGLOBIN(E) TECHNIC
HbA1c TECHNIC

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TROUBLESHOOTING THE CURVES
MINICAP
Please select a technique

PROTEIN(E) 6 TECHNIC
HEMOGLOBIN(E) TECHNIC

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TROUBLESHOOTING THE CURVES
MINICAP FLEX PIERCING
Please select a technique

PROTEIN(E) 6 TECHNIC
HEMOGLOBIN(E) TECHNIC
HbA1c TECHNIC

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 Protein(e) 6 Technic – Normal Control
MIGRATION PARAMETERS :
Temperature = 37.5 °C
Voltage = 7900 V (+/-300 V)
Current : 90 µA < I < 100 µA

PROTEIN – PAGE 1/17 – EXAMPLE/REFERENCE

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CURVE 1 : Protein(e) 6 Normal Control

88

PROTEIN – PAGE 2/17

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 CURVE 1 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Aged (old) capillary. - Aged (old) capillary.

The capillary cartridge must The capillary cartridge must
be replaced. be replaced.

- Bad cleaning of the

capillaries due to weak
shaker vibrations.
PROTEIN – PAGE 3/17
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CURVE 2 : Protein(e) 6 Normal Control

70

PROTEIN – PAGE 4/17

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 CURVE 2 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Bad activation - Bad activation
Renew the wash solution Renew the wash solution
using good quality of distilled water. using good quality of distilled water.

- Cathode reagent - Cathode reagent

Verify the correct draining Verify the correct draining
(from the cathode to the waste container) (from the cathode to the waste container)

(a message is displayed during the -Temperature regulation

priming in case of temperature Verify that the fan is correctly
regulation issue) pressed against the Peltier element.

PROTEIN – PAGE 5/17

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CURVE 2 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

MINICAP & F.P

-Temperature regulation
Verify that the fan is
correctly pressed against
the Peltier element.

PROTEIN – PAGE 5 Bis/17

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CURVE 2 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

IF UNITS WITH SN < 1750 MINICAP & F.P

-Temperature regulation
Verify that the fan is
correctly pressed against
the Peltier element.
IF UNITS WITH SN > 1750

Implemented on factory
since the SN (9)2812

PROTEIN – PAGE 5 Ter/17

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CURVE 3 : Protein(e) 6 Normal Control

88

PROTEIN – PAGE 6/17

Troubleshooting Curves - 2016 10 17
 CURVE 3 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

-Bad cleaning of the - Bad cleaning of the
capillaries : verify the capillaries due to weak
adjustments P261 and P265
shaker vibrations.

PROTEIN – PAGE 7/17

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CURVE 4: Protein(e) 6 Normal Control

PROTEIN – PAGE 8/17

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 CURVE 4 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Bad reagents (buffer and/or wash) - Bad reagents (buffer and/or wash)

- No dilution of serum (only air) - No dilution of serum (only air)

- Bad injection position - Bad injection position

(P261 too low) (P529 too low)

- Bad injection vacuum : syringe / - Bad injection vacuum :

EV18 / cathode tank / EV21 / EV5 syringe/8-way valve/cathode tank

- Optical issue : Lamp / - Optical issue : Lamp/filter wheel/

Monochromator / fibbers Emission optic fibbers Emission/capillary cell/
/ capillary cell / fibbers Recept. Optic fibber Recept.
PROTEIN – PAGE 9/17
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CURVE 5

PROTEIN – PAGE 10/17

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 CURVE 5 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Bad reagents (buffer and/or wash) - Bad reagents (buffer and/or wash)

- No dilution of serum (only air) - Bad dilution of serum (bad ratio)

- Bad injection position - Bad injection vacuum :

(P261 too low) syringe/8-way valve/cathode tank

- Bad injection vacuum : syringe / - Optical issue : Lamp / filter wheel /

EV18 / cathode tank / EV21 / EV5 optic fibbers / capillary cell

PROTEIN – PAGE 11/17

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CURVE 6

PROTEIN – PAGE 12/17

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 CURVE 6 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

Optical Noise Optical Noise

- Bad CCD calibration

- Faulty Lamp - Faulty Lamp

- Dirty optics fibbers - Dirty optics fibbers

(emission and/or reception) (emission and/or reception)

-Faulty Capillary -Faulty Capillary

(reading bubble) (reading bubble)

-Vibration of the optical fiber(s) -Vibration of the filter wheel

PROTEIN – PAGE 13/17

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CURVES 7

PROTEIN – PAGE 14/17

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 CURVES 7 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

Level of Buffer in the tanks. Level of Buffer in the tanks.

Cathode tank : Cathode tank :
- Sensor 4 - Air leak around the tank.
- Liquid Valve EV15 - Fluidics :
- Fluidic : Tubing Line 37 White Tubing Line
Syringe / 8-way valve

Anode tank : Anode tank :

- Sensor 3 - Syringe / 8-way valve
- Liquid Valves EV12 – EV13 - Needle / Green Tubing Line

PROTEIN – PAGE 15/17

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CURVE 8 (more difficult)

88

PROTEIN – PAGE 16/17

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 CURVE 8 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

Here, there is no problem Here, there is no problem

It’s a patient sample with an It’s a patient sample with an

example of a pathology example of a pathology
(gammopathy symptom) (gammopathy symptom)

MENU
PROTEIN – PAGE 17/17
Troubleshooting Curves - 2016 10 17
Hemoglobin technic – HbA2 normal Control

MIGRATION PARAMETERS :
Temperature = 36 °C
Voltage = 9650 V
Current : 80 µA < I < 100 µA

HB – PAGE 1/22 – EXAMPLE/REFERENCE

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HB – PAGE 2/22 – EXAMPLE/REFERENCE
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CURVE 1

No zone displayed

Position ≈ 200

FLAGS/ICONS RAW CURVE

HB – PAGE 3/22
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 CURVE 1 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

-Shifted migration speed -Shifted migration speed

Denatured Hb Buffer Denatured Hb Buffer

It remained at room It remained at room
temperature for more than temperature for more than
1 month (accumulated time). 1 month (accumulated time).

(a message is displayed during - Bad temperature regulation

the priming in case of (migration temperature is
temperature regulation issue) higher than the target)
HB – PAGE 4/22
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CURVE 1 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

HB – PAGE 5/22
Troubleshooting Curves - 2016 10 17
CURVE 2 : HbA2 normal control

FLAGS/ICONS RAW CURVE

HB – PAGE 6/22
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 CURVE 2 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Aged (old) control - Aged (old) control

After reconstitution, After reconstitution,

the HbA2 normal control has the HbA2 normal control has
been stored at 2°C-8°C for been stored at 2°C-8°C for
more than 1 week more than 1 week

HB – PAGE 7/22
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 CURVE 2 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

HB – PAGE 8/22
Troubleshooting Curves - 2016 10 17
CURVE 3 : HbA2 normal control

FLAGS/ICONS RAW CURVE

HB – PAGE 9/22
Troubleshooting Curves - 2016 10 17
 CURVE 3 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

Low Optical Density Low Optical Density

- Bad H2O volume of reconstitution: - Bad H2O volume of reconstitution:

(1ml /1.6 ml - Capillarys2 / F.P) (1ml /1.6 ml – MiniCap / F.P)

- Dilution issue (bad ratio) : syringe / - Dilution issue (bad ratio): syringe /
EV18 / tubing line / needle 8-way valve / tubing line / needle

- Bad injection position: P261 too low - Bad injection position: P529 too low

(Err. displayed if bad injection vaccum) - Bad injection vacuum :

syringe/8-way valve/cathode tank

- Optical issue : - Optical issue : Lamp / filter wheel /

Lamp / Optic Fibers / capillary cell optic fibers / capillary cell
HB – PAGE 10/22
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CURVE 4 : HbA2 normal control

FLAGS/ICONS RAW CURVE

HB – PAGE 11/22
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 CURVE 4 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

PLEASE REFER TO :

- LOW OPTICAL DENSITY

and

- USED HB BUFFER REAGENT

HB – PAGE 12/22
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CURVE 5 : HbA2 normal control

TOO LARGE

FLAGS/ICONS RAW CURVE

HB – PAGE 13/22
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 CURVE 5 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Bad cleaning of the - Bad cleaning of the

capillaries due to bad capillaries due to weak
adjustment : P261 and P265 shaker vibrations.

HB – PAGE 14/22
Troubleshooting Curves - 2016 10 17
CURVE 6 : HbA2 normal control

No zone displayed

- Wasted buffer

- No temperature regulation Position ≈ 175

Position ≈ 100

FLAGS/ICONS RAW CURVE

HB – PAGE 15/22
Troubleshooting Curves - 2016 10 17
 CURVE 6 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Aged (old) capillary - Aged (old) capillary

In this case : In this case :

The O.D is increased The O.D is increased

The curve is shifted to the The curve is shifted to the

right hand side. right hand side.

 Replace the capillary  Replace the capillary

HB – PAGE 16/22
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CURVE 7 : HbA2 normal control

No zone displayed

- Wasted buffer

- No temperature regulation
Position ≈ 100

FLAGS/ICONS RAW CURVE

HB – PAGE 17/22
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 CURVE 7 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Cold Hb Buffer: it must slowly returns - Cold Hb Buffer: it must slowly returns
to room temperature before to be used. to room temperature before to be used.

- Bad activation : renew the Wash Sol. - Bad activation : renew the Wash Sol.

(Err. Message in case of - Temperature regulation issue

temperature regulation issue) (migration temperature is lower
than the target)

- Cathode reagent - Cathode reagent

Verify the correct draining Verify the correct draining
(from the cathode to the waste container) (from the cathode to the waste container)

HB – PAGE 18/22
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CURVE 8 : HbA2 normal control

No zone displayed

- Wasted buffer

- No temperature regulation
Position ≈ 100

FLAGS/ICONS RAW CURVE

HB – PAGE 19/22
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 CURVE 8 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

Optical Noise Optical Noise

- Faulty Lamp (light is not stable) - Faulty Lamp (light is not stable)

-Vibrations : -Vibrations on the filter wheel

(monochromator and/or optical fiber(s))
- Bad position of the filter wheel
 Low optical levels  optical noise

- Faulty capillary (reading window) - Faulty capillary (reading window)

HB – PAGE 20/22
Troubleshooting Curves - 2016 10 17
 FLAGS / ICONS

HbA and/or HbA2 are not detected inside the expected target window
(fractions are out of zones)

The recentering of the curve is not possible

Optical density of the normal HbA2 control is too low

MENU
HB – PAGE 21/22
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CURVE(S) : VISUALIZE THE RAW CURVE

MENU
HB – PAGE 22/22
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HbA1c Technic – Calibrator (level1 or level 2)

MIGRATION PARAMETERS :
Temperature = 36 °C
Voltage = 9200 V
Current : 70 µA < I < 80 µA

HbA1c – PAGE 1/19 – EXAMPLE/REFERENCE

Troubleshooting Curves - 2016 10 17
HbA1c Technic – Control (level 1 or level 2)

MIGRATION PARAMETERS :
Temperature = 36 °C
Voltage = 9200 V
Current : 70 µA < I < 80 µA

HbA1c – PAGE 2/19 – EXAMPLE/REFRENCE

Troubleshooting Curves - 2016 10 17
CURVE 1

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 3/19
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 CURVE 1 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

- Aged (old) capillary - Aged (old) capillary

In this case : In this case :

The O.D is increased The O.D is increased

The curve is shifted to the The curve is shifted to the

right hand side. right hand side.

 Replace the capillary  Replace the capillary

HbA1c – PAGE 4/19

Troubleshooting Curves - 2016 10 17
CURVE 2

Position ≈ 180

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 5/19
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 CURVE 2 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

Shifted migration speed Shifted migration speed

- Wasted HbA1c Buffer -Wasted HbA1c Buffer

More rare : More rare :

- Migration temperature increased -Migration temperature increased
- Migration High Voltage increased -Migration High Voltage increased
(issue after a change of technic or (issue after a change of technic or
corrupted parameter) corrupted parameter)
HbA1c – PAGE 6/19
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CURVE 3

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 7/19
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 CURVE 3 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

OPTICAL NOISE OPTICAL NOISE

Vibration on the optics fibers

(emission and/or reception)

Faulty capillary reading cell Faulty capillary reading cell

Monochromator : bad vertical Filter wheel : bad position

position and / or vibrations and / or vibrations

HbA1c – PAGE 8/19

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CURVE 4

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 9/19
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 CURVE 4 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

. Optical Noise : . Optical Noise :

Vibration on the optics fibers Vibration on the filter wheel
(emission and/or reception)

. Potential shifted migration : . Potential shifted migration :

The baseline is not correctly The baseline is not correctly
placed by the software placed by the software

HbA1c – PAGE 10/19

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CURVE 5

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 11/19
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 CURVE 5 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

OPTICAL NOISE OPTICAL NOISE

Please defocus the emission bundle

Faulty capillary reading cell

Filter wheel : bad position

and / or vibrations

HbA1c – PAGE 12/19

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 CURVE 5 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P

No more noise : the defocusing is fine

HbA1c – PAGE 13/19

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CURVE 6

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 14/19
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 CURVE 6 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P

• Low Optical Density :
Bad reconstitution volume
(the correct volume is 0.6 ml of distilled water)

Bad injection vacuum (between syringe and vacuum sensor)

• Curve Position :

Bad activation (Wash Solution)

Cold HbA1c Buffer

High Voltage decreased (issue after a change of technic)

HbA1c – PAGE 15/19

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CURVE 6 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

MINICAP & F.P

• Low Optical Density :
Bad reconstitution volume
(the correct is 0.6 ml of distilled water)
Bad injection vacuum (between syringe and cathode tank)

• Curve Position :
Bad activation (Wash Solution)
Cold HbA1c Buffer
Migration Temperature decreased
High Voltage decreased (issue after a change of technic)
HbA1c – PAGE 15/19
Troubleshooting Curves - 2016 10 17
CURVE 7

FLAGS/ICONS RAW CURVE

HbA1c – PAGE 16/19
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 CURVE 7 : POTENTIALS CAUSES - POTENTIALS ITEMS INVOLVED

CAPILLARYS 2 & F.P MINICAP & F.P

• No cleaning of the capillaries • No cleaning of the capillaries

due to motor2 out of order. due to a shaker
Message is displayed out of order.
during the priming cycle. Replace the shaker

• Unstable level of Buffer • Unstable level of Buffer

inside the cathode tank during inside the cathode tank during
the migration the migration
 EV15 is involved  EV and/or 8-way valve are
involved.
HbA1c – PAGE 17/19
Troubleshooting Curves - 2016 10 17
 FLAGS / ICONS

HbA1c – PAGE 18/19

Troubleshooting Curves - 2016 10 17 MENU
CURVE(S) : VISUALIZE THE RAW CURVE

MENU
HbA1c – PAGE 19/19
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 HOW TO APPROACH ”BAD” CURVES ?
1 - Is it a recurrent problem or an isolated case?
All the capillaries are affected, or is it only one position ?
 if only one position is affected, then a dilution issue can be suspected.
Rerun this sample into another capillary position to prevent an issue with a specific capillary position

 If all the positions are affected, then we can suspect a common item :
Reagents : Wash solution and/or Buffer / and/or diluant(s)
Optics : Lamp, wave length, reception PC Board
Fluidics : injection vacuum, Buffer level on the tank(s)
Mechanics : injection position, cleaning position

2 - Does it happen only for a specific technique ?

 If so, then we can suspect an issue with the Buffer or during the change of technique

3 – What has changed, what has been modified, just before the phenomenon ?

→ The same type of strange curves can be due to different potentials causes : insufficient volume of sample, presence of an air
bubble in the tubing line, optical issue, mechanical adjustments… etc.

→ If a repeating problem is suspected, looking at one curve only is not sufficient.

All the curves of this working day must be analyzed…
And sometimes several days, for several techniques.

MENU REMINDER Troubleshooting Curves - 2016 10 17

 REMINDER
• All the Technics :
Migration too slow (shifted to the left) : potential bad Wash Solution or bad activation
Migration too slow (shifted to the left) : potential mix between Buffer and Wash Solution
Migration too fast (shifted to the right): potential mix between Buffer and Water

• Migration Parameters :
The speed of the migration (i.e the curve position) depends on :
The Reagents - The migration High Voltage - The migration temperature.

The O.D (amplitude of the curve) depends on :

The injection vacuum – The Optical levels – The cleaning of the capillaries

• Hb and HbA1c Technics :

A cold Buffer will slow down the migration : the curve is shifted to the left
A wasted or aged Buffer (*) will accelerates the migration : the curve is shifted to the right
(*) because at room temperature since too long
MENU Troubleshooting Curves - 2016 10 17
 TROUBLESHOOTING THE CURVES

THE END

MENU Troubleshooting Curves - 2016 10 17