Capillarys 2 & Capillarys2 Flex Piercing

How to troubleshoot a dilution issue

Table of contents
Introduction ................................................................................................................................................................... 2

Running samples ......................................................................................................................................................... 2

DCN calibration ............................................................................................................................................................ 3

Mechanical adjustments ............................................................................................................................................. 4

Most common parts to replace .................................................................................................................................. 8

Appendix1 : Test Cycle C16 ...................................................................................................................................... 11

Appendix2 : Mechanical play of the syringe ........................................................................................................... 12

Appendix3 : Capillarys2 Flex Piercing – Draining line .......................................................................................... 14

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 Introduction :
Before any action, or parts replacement, it’s necessary to observe very carefully the dilution process.
The level (and the color) inside each comb of the dilution segment is a first indication :

Note :
- insufficient volume of serum or blood will generate a low O.D
- insufficient volume of Hemolysing Solution will generate a high O.D

To highlight a dilution issue, please run a SEBIA rack with a segment manually diluted :

Volume
aspirated
Serum / Blood Buffer Diluant/Hemolysing Water
Technique
Protein(e) 6 40 µL 160 µL not used not used
C.D.T 40 µL not used 40 µL 120 µL
Hemoglobin(e) 18 µL not used 90 µL not used
HbA1c 18 µL not used 90 µL not used

- If the issue is fixed when running the SEBIA rack, then we are facing a dilution problem.
It could be related to the mechanical play of the syringe (see Appendix2).

- If the issue is not fixed, we are facing another issue (capillaries, injection vacuum, optics, …etc).

Running samples :

 Make sure there is a good vacuum inside the wash station (no dripping underneath).
In case of dripping, it could be a leak from the waste container, a dirty air filter, a weak pump
(make the pump PM), or even an issue with a liquid valve.
Verify that the needle is correctly positioned inside the wash station

 Make sure there is no air bubble inside the syringe

(replace the syringe seal and piston if necessary).
The valve EV18 could be involved : bad calibration or bad mechanical coupling.
All the tubing lines connected to EV18 must be 100% full of liquid.

 Verify that the needle is correctly positioned upon the sample tubes and upon the dilution
segment. Refer to the mechanical calibrations (hereafter).

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 DCN calibration :
Preferably, it should be done when the instrument is running Protein(e) 6 technic.

It's mandatory to use the correct tube and the correct

Eppendorf micro-tube to perform the DCN calibration.
Not a shorter or different shape of micro-tube

Before renewing the DCN Calibration, perform a

Capiclean cycle in order to clean the needle.
The Capiclean cycle being mandatory once a week
(customer’s maintenance).

Expected range : 400 < Threshold < 800 4000 < Offset < 8000

Exceptionally when the detection is too sensitive (probe sensing something in the air before reaching
the serum), we can make a DCN calibration with 200 µl of distilled water instead of 100 µl into the
Eppendorf tube. It will decrease the sensitivity of the needle.

Heigh of the Needle Calibration : preferably, it should be done using the Protein(e) 6 technic.

Now, an empty IT segment is provided with the instruments. Fill the double comb with 380 µL of
distilled water and then place this segment the Maintenance rack (#100).
Launch the maintenance rack and select “Calibration of the sample probe”.

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 Mechanical adjustments :

Check the mechanical coupling of the

motor 5 (up/down) and motor 6 (backward/forward).
Lubricate the endless screws using
the white mechanical grease
(after removing the old grease).

Lubricate the 4 axis using liquid silicon oil

(after cleaning the axis).

 Make sure the needle is well calibrated in the 3 directions :

- left/right with the customized parameter P51
- forward/backward with the customized parameters P69 & P274.

The needle must be centered on the middle of the sample tube

(for the pipetting) and centered on the segment (for the dilution).

 Make sure the extremity of the needle is correctly positioned inside the wash station.
From the Ready mode, adjust the sample probe extremity in the middle of the wash station with the
help of the 2 Allen screws on the top of the sample probe.

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 Make also sure that the head syringe housing is mounted into the correct direction : the internal
channel is into the upper side. The brown coupling must not be over tightened (pinched).

 Verify the correct electrical continuity of the yellow wire (no bad contact on the top of the needle)

The extremity of the needle must be in

continuity with the yellow cable

plugged into the DCN PC Board :

 connector J5 (pin 1)

 or center conductor of connector J3

Note :
the external wall of the needle is Isolated.

Only the exact extremity and the inner wall

is an electrical conductor

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  Check the electrical continuity all along the shielded sample tubing.
Starting from the black wire (ground) on the DCN PC Board until the end.

See pictures : Point A must be an equipotential with Point B…even when

the needle is moving up/down

An internal bad contact of the shielded cable is hardly visible, so it can be good to replace it.

 Check all the grounding connections inside the instrument. Make sure the earth is good on the wall
socket, plug the power cord to another AC wall socket to eliminate any potential issue on this side.

 The opto sensor (bubble sensor) located behind the needle assembly is not used.

So please remove the tubing line from this sensor :

doing so, the cable is totally free to move during
the needle’s movement up / down,
without damaging the black wire.

But, please note that the shielded sample tubing

is isolated from the chassis
(ground of the instrument).

The shielded sample tubing must never

touch any metallic part
(chassis or syringe’s motor)

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 Verify that there is no air leak between the syringe and needle

 The wash station of the needle must be clean and not obstructed. Otherwise, there is a bad
cleaning of the needle and it could remains wet when using the DCN sensitivity.

The cleaning is correct if there is a good aspiration (vacuum) inside the wash station.
The waste tubing line must never be pinched or obstructed. All along the way between the wash
station and the waste container

It’s the exact same principle on Capillarys2 Flex Piercing instruments, except that there are 2 liquid
valves on the wash station : EV4 and EV6.
The 4-way valve EV18 is replaced by a 6-way valve (or 8-way valve) EV18.
The liquid valve EV7 is replaced by a pinch valve EV7.
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Make also sure that there is no air leaks on the waste container.

Most common parts to replace :

- Syringe O’ring : P/N 40000262

- Sample probe (internal coating scratched, yellow wire broken) : P/N 10051212
(Capillarys 2 Flex Piercing : P/N 31003369E)
The beveled needle can be unblocked using a piano string :
diameter 0.3 or 0.4 mm – length 20 cm (around 8 inches)

- Pump maintenance kit : P/N 10051904

- 4 way valve (EV18) : P/N 10051902
(8-way valve on Capillarys 2 Flex Piercing : P/N10052720)
(6-way valve on Capillarys 2 Flex Piercing : P/N40000607)
- Needle tubing (shielded cable + tubing pipe) : P/N 41000008A
(Capillarys 2 Flex Piercing : P/N 41000015A)

- Pinch Valve on Capillarys2 Flex Piercing (EV17) : P/N 45360036

- Pinch Valve tubing line on Capillarys2 Flex Piercing : P/N : 40000140 (sold by the meter).

Note that the DCN PC board (P/N 10046223) is almost never responsible of a detection issue.

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 Troubleshooting with the service menu : Test Cycle C16 - See Appendix1

This cycle is not applicable on the releases 6.50, 6.51 and 8.50 (the graphs obtained are not usable -
significant). It can be performed on the releases 6.12, 8.51 and 8.60 and next.

Please note that when running Phoresis Core (release 8.60), the priming of the tubing line (after a
change of technic) is now improved, to avoid missing the first dilutions.

Using the test cycle C16 we can visualize the DCN level detection of the needle during a dilution.

When the needle touches the liquid, the signal detected by the needle is modified.
The more liquid there is on the tube, the more the difference of level (Threshold) will be important.
In our case (please refer to the Appendix1), the quantity of distilled water in the tubes is descending.
So, the threshold of the signal returned by the needle is more and more weak.

Place 8 sample tubes on a rack. Each tube contains

a volume of distilled water (from 5 ml to 100µl).

Refer to the Appendix1.

For the dilution positions containing less than 1ml,

please put the distilled water into Eppendorf micro tubes.

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 Examples of DCN issues when running C16

Check the correct grounding connections and the welding spots on the shielded cable

-----------------------------------------------------------------------------------------------------------

The needle seems to be already

wet before touching the liquid.

EV17 and the vacuum line

(between the wash station and
the waste container) are very
probably involved

-----------------------------------------------------------------------------------------------------------

Internal liquid leak of EV18

-----------------------------------------------------------------------------------------------------------

The shielded sample tubing is touching a

metallic part (chassis or syringe’s motor).

Or the yellow DCN cable of the needle is

cut, or poorly welded (bad soldering point).

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 Appendix1

Using the test cycle C16 we can visualize the DCN level detection of the needle during a dilution
process. When the needle touches the liquid (serum or H2O), the signal detected by the needle is
modified. The more liquid there is on the tube, the more the difference of level will be.

In our case (attached pictures), the quantity of distilled water in the tubes is descending.
So, the threshold observed is increasingly weak.

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 Appendix2

 Mechanical play of the syringe :

This mechanical play is measured only during

the very first priming of the day.

So, in case of replacement of the syringe,

it’s necessary to select the option “Force complete start”.

The measurement is done after the initialization of the motors :

1st step : syringe on the home position
2sd step : syringe moves 100 µsteps
3rd step : syringe comes back to home position in order to compares the µsteps done

The mechanical play indicates the correct functioning of the syringe (for the dilution ratio).
This value is saved on the EEPROM, at the address 400 (Length = 01).

When the instrument is on « Ready », reach the EEPROM window.

Enter the address 400 and click on Read. The value displayed is an Hexadecimal value (Hex).

The mechanical play shall be less than 64 (Hexa value). So 100 motor steps (decimal value).

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It’s possible to force the measured value using the parameter P298 :

If this value is forced (so different from 0), the measured value (first priming of the day) is not taken
into account. Only the forced value is applied.

Example : dilution issue, visually confirmed with different levels of dilution inside the dilution
segment. In such a case, read the measured mechanical play (address 400), and then force it
with P298 but enter a forced value lower than the measured one (10 steps less should be enough).

Important :
with time the mechanical play will increase, so the replacement of the piston will be necessary.

------------------------------------------

Convert Hexa to Decimal :

The base hexadecimal is a numeral system with a base of 16 (using 16 different symbols)
The base decimal is a numeral system with a base of 10 (using 10 different symbols)

We must decompose each hex character, then multiply by 16 N

(N being the weight value of the bit  0 for LSB*).
Each hex character is converted to a decimal value. Finally add the decimal values.

Example : convert the hexa value 04F

Here, « F » is the LSB (0), and «0 » is the MSB (2 in this case).

Hexa Décimal  04F (Hex value) is 0 + 64 + 15 = 79 (decimal value)

0 0x162=0

4 4x161 = 64

F Fx160 = 15x160 = 15x1 = 15 * LSB : Least Significant Bit - MSB : Most Significant Bit

------------------------------------------

Calculation :
0
16 =1
1
16 =16x1=16
2
16 =16x16=256

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 Appendix3
Verify the draining tubing line on Capillarys2 Flex Piercing instruments

The draining line goes from the wash station of the needle to the external waste container.
Reminder : on the couplings of the waste container, any small air leak will generate a bad draining and
therefore a bad cleaning of the sample probe. Consequently the dilution cycle is disturbed.

In some locations (all along this draining line), a tubing could gets pinched, or obstructed.

It’s possible to modify the routing of the tubing lines, in order to prevent any pinching or plugging :

Verify that the tubing inside the pinch valve

EV17 is not distorted  always closed.
Do not hesitate to remove this tubing line,
shift it from 1 cm and then place it back
inside the pinch valve EV17.
If necessary, this tubing line can be
provided as a spare part with the part
number 40000140 (unit : 1 meter).

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 BY DEFAULT

The draining tubing line is placed upon the metal

chute of the reagent.

POSSIBLE MODIFICATION

OR

The draining tubing line could be placed

underneath the metal chute of the reagents. The draining tubing line could be placed lower than the metal
The tubing line would be maintained by 2 chute of the reagents. The tubing line would be maintained
colrings, attached on the vertical bars. by a holder, glued on the low voltage power supply.

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