SYSMEX CyStain VitalCount User manual

CyStain™ VitalCount | Quality Check with CyView™ 1.8
Doc. No.: 7 QC IFU EN | Rev: 001 | Date: 25-06-2021 | EN | CN 2138
CyStain™ VitalCount reagents
Quality Check Manual

Table of contents
2 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
Table of contents
1Identification ..............................................................................................................3
2Introduction................................................................................................................4
2.1About this document......................................................................................4
2.2About the procedure.......................................................................................4
2.3Installation and alteration ..............................................................................4
2.4Sysmex representatives.................................................................................4
2.5Authorised distributors..................................................................................4
3Overview.....................................................................................................................5
3.1CyView™script...............................................................................................5
3.2FCS Express™ template ................................................................................5
3.3Quality Check material ...................................................................................5
3.4QC material LOT dependency........................................................................6
4Operation....................................................................................................................7
4.1Software...........................................................................................................7
4.2Quality Check procedure ...............................................................................7
4.2.1Setting up the Quality Check ............................................................................ 7
4.2.2Running a Quality Check .................................................................................. 8
4.2.3Analysing the Quality Check with FCS Express™.......................................... 11
4.3Measurement procedure ..............................................................................12
5Troubleshooting ......................................................................................................13
5.1Fault, cause and remedy..............................................................................13
5.1.1Sheath Fluid, waste and fluids ........................................................................ 13
5.1.2QC Procedure ................................................................................................. 14

1 Identification
CyStain™ VitalCount QC Procedure | QC Manual | June 2021 3
1 Identification
The Quality Check procedure is applicable to:
Reagent / Reagent kit
Name CyStain™ VitalCount
Manufacturer Sysmex Partec GmbH
REF 05-5029
Device
Name CyFlow™ Cube 6 V2m
Software CyView™ 1.8 and above
Manufacturer Sysmex Partec GmbH
REF CY-S-3061R-V2m

2 Introduction
4 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
2 Introduction
2.1 About this document
This document will guide through the Quality Check (QC) procedure of the CyStain™
VitalCount reagent kits with a CyFlow™ Cube 6 V2m. Emphasis is placed on execution and
troubleshooting.
Please refer to the CyFlow™ Cube 6 V2m manual for correct and safe use of the device.
For questions about this manual or the use of the device, please contact your local Sysmex
representative.
2.2 About the procedure
The Quality Check scripts simplify and automate the QC procedure of the CyFlow™ Cube
6 V2m for measurements with the CyStain™ VitalCount reagent kits. For analysis of the
results, they are combined with associated FCS Express™ templates that are automatically
launched during the QC procedure. It is necessary to apply specific Quality Check scripts
and FCS Express™ templates corresponding to the used QC material LOT.
2.3 Installation and alteration
The first installation must be performed by authorised service personnel.
No alterations should be made to the scripts and templates, unless expressly permitted by
the manufacturer. After alteration of scripts or templates appropriate tests and trials must
be performed to ensure safe use.
2.4 Sysmex representatives
Find address and contact data of your local Sysmex representative at
www.sysmex-flowcytometry.com
2.5 Authorised distributors
To receive an overview of authorised distributors nearby, please contact your local Sysmex
representative.

3 Overview
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3 Overview
3.1 CyView™script
The Quality Check procedure is integrated into a CyView™ configuration file (*.cvc85) -
called a CyView™ script. To access the QC procedure, the “QC”-Mode within the respective
CyView™ script can be activated.
3.2 FCS Express™ template
FCS Express™ templates are used to automatically analyse the results of QC
measurements. The templates can also be used to analyse data from QC measurements
manually. The results can be exported.
3.3 Quality Check material
The QC procedure uses the following material(s). For further information please refer to the
device manual.
NOTICE
Count Check Beads green specificity
This QC procedure was designed for the use with Count Check Beads
green (REF 05-4011_R). Similar products such as Count Check Beads
green (REF 05-4011) are not suitable for use.
Always use the recommended reagents.
Light source Item Content REF
Blue Laser Count Check Beads green 50 tests 05-4011_R

3 Overview
6 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
3.4 QC material LOT dependency
The CyView™ script as well as the FCS Express™ templates are designed for a specific
Count Check Beads green (05-4011_R) LOT.
Check before QC procedures
LOT of used Count Check Beads green and revision of the script / templates are matching
(see 4.2 Quality Check procedure).
“LOT N° mismatch” will be displayed in the FCS Express™ templates, if a template not
matching the Count Check Beads green LOT was used.
The Quality Check is designed solely for QC material stated in chapter 3.3 Quality Check
material. Similar products, such as Count Check Beads green (REF 05-4011) are not
suitable.
Matching LOT and template
The LOT number (LOT No) of the Count Check Beads green is composed of
“NN00YYMMDD”. They are compatible with the script / template version, identical to the
first 4 digits of the LOT No“NN00”.
To check if the LOT matches, start the FCS Express™ template manually and type in the
LOT Noof the Count Check beads green. If the LOT Nodoes not match “LOT N° mismatch”
will be displayed.
Find the template at: “C:\ProgramData\PartecGmbH\Cube_18\templates\Quality Control”

4 Operation
CyStain™ VitalCount QC Procedure | QC Manual | June 2021 7
4 Operation
4.1 Software
CyView™ is used for control of the device, data acquisition, data analysis and storage. For
more information, please refer to the CyFlow™ Cube 6 V2m manual.
FCS Express™ is used for reporting and analysis of data. The software can be accessed
directly from within CyView™. Refer to FCS Express™ manuals and tutorials for detailed
information about the software.
4.2 Quality Check procedure
4.2.1 Setting up the Quality Check
Requirements
Logged in to CyView™ with privileges of Main User level or above (Administrator)
LOT and FCS Express™ template match, see chapter 3.4 QC material LOT
dependency
CyStain™ VitalCount configuration is active1.
Priming has been performed
Sample tube with 850 μl Count Check Beads green, see chapter 3.3 Quality Check
material
Sample tube with 1500 μl Sheath Fluid
Procedure
Figure 6:Selecting Work Mode
1. Select [Work Mode].
2. Switch Measure Mode Sample Source to “Sample Port”.
3. Connect the Sample tube with 850 µl Count Check Beads green.
4. Click [Start].
5. Adjust gain values in a way that the signals of the Count Check Beads green appear
exactly in the center of the quality gate “QC (CCB)” within the main Plot.
DQC Plot 1 DQC Plot 2
Figure 7:Adjust gain values
1E.g. VitalCount_A_AK01_Robby.cvc85

4 Operation
8 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
6. Click [Stop].
7. Connect the Sample tube with 1500 μl Sheath Fluid.
Figure 8: Intermediate Cleaning
8. Activate [Intermediate Cleaning].
9. Switch Measure Mode (Sample Source) back to “Autoloader”.
10. Switch Measure Mode (Sample Volume) to “51 – 100 µl”.
11. Switch Measure Mode (Sample Mixing) to “High”.
12. Switch Measure Mode (Sample-to-Sample Cleaning) to ”Intensive”.
13. Switch back to “Prime Mode” and save the configuration file as …_Robby.
Result
The configuration file containing suitable gain values for all parameters can be saved on
the device. It is recommended to maintain the default name for the configuration file.
Continue by running a Quality Check.
4.2.2 Running a Quality Check
The QC procedure controls background, laser power, optical alignment, gate positioning
and counting precision.
Requirements
Logged in to CyView™ with privileges of Main User or Administrator
LOT and FCS Express™ template match, see chapter 3.4 QC material LOT
dependency
CyStain™ VitalCount configuration is active
Priming has been performed
96-Well Plate is selected with QC material in the following order (e.g. A1)
- e.g. A1 well with 200 μl Count Check Beads green

4 Operation
CyStain™ VitalCount QC Procedure | QC Manual | June 2021 9
Procedure
Figure 9:Selecting QC Robby Mode
1. Select [QC Robby Mode].
2. Press [Start] and follow the instructions.
3. The CyView™ Robby Autoloader AddOn will open. Assign a new Tray ID, e.g.
“QCyyyymmdd”.
Figure 10:CyView AddOn
4. Click [Start with new Tray].
5. Select each with QC material and insert the Wellname “QC”. The correct order for
the QC material is important when measuring the wells.
6. Click “Back to CyView”.
Figure 11:CyView TrayLayout

4 Operation
10 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
7. Press [Start] again to run QC Robby.
Figure 12:CyView Start Robby
8. Follow the instructions and Click [Continue] to start FCS Express.
NOTICE
Adjusting Device Settings during a Quality Check
Signals should be visible in all dotplots. The predefined QC regions
indicate the position where the signals of the beads are expected.
If the beads signals are not within the predefined region, adjust the gain
values , see chapter 4.2.1 Setting up the Quality Check.
Results
The QC procedure results can be analysed in FCS Express™. Continue by analysing the
Quality Check with FCS Express™.

4 Operation
CyStain™ VitalCount QC Procedure | QC Manual | June 2021 11
4.2.3 Analysing the Quality Check with FCS Express™
FCS Express™ opens automatically, once the QC procedure has finished. The QC results
will be analysed automatically by FCS Express™.
The first 4 digits of the LOT number need to be entered (e.g. “AK01”) in the first part of the
LOT N° field. Make sure that the field contains no further characters, such as spaces.
Figure 13: Exemplary QC report (Page 1 / 2)
(before Enter Quality Check material information) (after Enter Quality Check material information)
# Description # Description
1 Input field Quality Check material information:
LOT Number (example “AK01”)
5 Summary of performed tests Test: Pass or Fail
2 Input field Quality Check material information:
Expiry Date
6 General information Device serial number,
CyView Operator, Sample ID and Date
3 Input field Quality Check material information:
Reference concentration
7 FCS Express™ template version
4 QC report summary: Device status: Valid or
Invalid
1
2
3
4
5
6
7

4 Operation
12 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
Procedure
1. Switch to FCS Express™. Data have been transferred automatically to a predefined
Layout.
2. Enter Quality Check material information for LOT No, expiry date and reference
concentration for Count Check Beads green.
3. The QC results are analysed automatically. Every test performed will “Pass” or “Fail”
and a summary will be displayed.
4. To save a .pdf document of the results, switch to the Batch tab within FCS
Express™ and click [Run]. Choose a suitable file folder and assign a data name. A
PDF report will be generated and saved into the selected folder.
Results
If all tests are passed, the device is qualified to perform a measurement with the reagent
kit. If one or more tests fail, the device is not qualified to perform the measurement. Please
refer to chapter 5 Troubleshooting.
4.3 Measurement procedure
Please refer to the CyStain™ VitalCount kit manual as well as to the CyFlow™ Cube 6 V2m
manual for instructions on perfoming measurements, saving aquired data as well as
cleaning and shut down procedures.

5 Troubleshooting
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5 Troubleshooting
5.1 Fault, cause and remedy
If the fault you are experiencing is not described or the remedy could not solve your
problem, please contact your local Sysmex representative.
NOTICE
User Privileges
Options for troubleshooting depend on the user privileges. Controls for
lasers, gain, flow rate and trigger parameter are only available at Main User
level or above.
5.1.1 Sheath Fluid, waste and fluids
Fault Remedy
Sample flow is very slow or
is not running
Check the flow rate settings.
Check that the Waste bottle is closed properly and that the
bottle is not cracked.
Check all visible tubes and make sure that they are not
pinched.
Make sure that no air bubbles are trapped in the Inline
Filter of the Sheath Fluid bottle.
Perform priming procedure.
Perform cleaning procedure.
Shut down the device, including the Power switch on the
back side of the device. Wait 3-5 minutes and restart the
system.
Sample flow is very fast even
with low flow rate
Check the speed settings.
Change the Inline Filter in the Sheath Fluid bottle.
Check if the visible Sheath Fluid tubing is pinched or
blocked.
Shut down the device, including the Power switch on the
back of the device. Wait 3-5 minutes and re-start the
system.

5 Troubleshooting
14 CyStain™ VitalCount QC Procedure | QC Manual | June 2021
Fault Remedy
The device emits an unusual
/ noisy sound after starting
the measurement
Check that the Waste bottle is closed properly and that the
bottle is not cracked.
Check all visible tubing and make sure that they are not
pinched.
Check that liquid (Sheath Fluid and sample) is dropping
into the Waste bottle after starting the measurement.
5.1.2 QC Procedure
Fault Remedy
There is no data acquisition
visible (peaks/dots in
histogram/dot plot) during
the RUN phase
Check within the software if the laser is switched on.
Check if the right trigger parameter (FL1) has been
selected.
Check and adjust Gain values for all parameters
Check threshold.
Unusual high background
during measurement and /or
failling background test
within the Quality Check
report
Make sure that no air bubbles are trapped in the Inline
Filter of the Sheath Fluid bottle.
Check threshold.
Perform a priming procedure.
Perform a cleaning procedure.
Refill the Sheath Fluid. Sheath Fluid should be refilled
every day.
Clean the Sheath Fluid bottle and rinse it with Hypochlorite
Solution.
Renew the Inline Filter. The Inline Filter should be
replaced every 3 months.
Check the date of expiry of the QC Material.
1. Remove the Inline Filter.
2. Fill the Sheath Fluid bottle with Hypochlorite Solution
and reconnect it.
3. Switch to [Analyze All] and run three times 1200 µl
Hypochlorite Solution at full speed.
4. Refill the Sheath Fluid bottle with fresh Sheath Fluid
and run three times 1200 µl Sheath fluid at full speed.
Failling Laser Power test
within the Quality Check
report
Check within the software if the laser is switched on.
Check and adjust gain values for all parameters.
Check the date of expiry of the QC material.
Shut down the device, including the Power switch on the
back side of the device. Wait 3-5 minutes and re-start the
system.

5 Troubleshooting
CyStain™ VitalCount QC Procedure | QC Manual | June 2021 15
Fault Remedy
Unusal large CV values
and/or failling Optical
Alignment test within the
Quality Check report
Perform a priming procedure.
Perform a cleaning procedure.
Check the date of expiry of the QC material.
Shake the bottle of the QC material vigorously (e.g. by
vortexing) and repeat the measurement.
Check and adjust gain values for all parameters.
Count Check Beads green
do not appear in the
predefined regions of the
configuration script and /or
failing Gate Position test
within the Quality Check
report
Check within the software if you have loaded the correct
configuration file for your measurement.
Shake the bottle vigorously (e.g. by vortexing) and repeat
the measurement.
Check the date of expiry of the QC material.
Perform a priming procedure.
Perform a cleaning procedure.
Adjust gain values for all parameters
Count Check Beads green
measurement is not within
± 20 % range of the LOT-
specific concentration stated
on the bottle and /or failing
Counting test within the
Quality Check report
Shake the bottle vigorously (e.g. by vortexing) and repeat
the measurement.
Check the date of expiry of the Count Check Beads green.
Perform a priming procedure.
Perform a cleaning procedure.
Refill the Sheath Fluid (ideally daily).
Renew the Inline Filter. The Inline Filter should be
replaced every 3 months.
Check and adjust Gain values for all parameters.
“LOT N° mismatch” is
displayed in the “Device
Status” after entering the QC
Material LOT number
Make sure to use a matching FCS Express™ template
and Count Check Beads green LOT.
The LOT number of the Count Check Beads green is
composed of “NN00YYMMDD”. They are compatible with
script / template version identical to the first 4 digits of the
LOT number “NN00”.