SYSMEX CyFlow Space User manual

For Research Use Only. Not for use in diagnostic procedures.
With FloMax®2.1x software
Doc. No.: CY-S-3001R-FM2IFUEN | Rev.: 013 | Rev. date: 18-03-2021 | EN | CN 391
CyFlow
™Space
Operating Manual

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Table of contents
1Identification............................................................................................................5
1.1Product information .........................................................................................5
1.2Manufacturer.....................................................................................................5
2Introduction.............................................................................................................6
2.1What is the CyFlow™ Space? .........................................................................6
2.2What are the applications for which the CyFlow™ Space can be used?....6
2.3What topics are covered by this manual?......................................................6
2.4What other manuals are available?.................................................................6
2.5What should I know before operating the CyFlow™ Space?.......................6
2.6What standards were applied for CE compliance?.......................................7
2.7What typographical conventions are used in this manual?.........................7
2.7.1Symbols in this document...................................................................................7
2.7.2Safety information...............................................................................................7
3Typical Steps of Particle Analysis.........................................................................9
3.1Preparation and Staining.................................................................................9
3.2Flow Cytometry Analysis.................................................................................9
3.3Real-time Data Processing and Results.........................................................9
3.4Absolute Cell Counting..................................................................................10
4Safety .....................................................................................................................11
4.1Laser radiation hazards .................................................................................11
4.2Electrical hazards...........................................................................................11
4.3Alterations to the device................................................................................11
5Operating Basics...................................................................................................12
5.1Switching on the CyFlow™Space ................................................................12
5.1.1Check SHEATH and WASTE bottle .................................................................12
5.1.2Switch on the instrument ..................................................................................12
5.1.3Control of the 488 nm laser output power (for 200 mW high power laser only,
model Coherent Sapphire)...........................................................................................12
5.1.4Switch on peripheral devices............................................................................12
5.1.5Switch on computer..........................................................................................12
5.1.6Start instrument operating software..................................................................13
5.1.7Priming the instrument (initial cleaning)............................................................13
5.2Multi laser Measurements..............................................................................13
5.3Starting a Measurement.................................................................................14
5.4Switching off the CyFlow™ Space................................................................16
6True Volumetric Absolute Counting – Overview................................................17
6.1Non-volumetric Flow Cytometric Techniques for Absolute Counting ......17
6.2True Volumetric Absolute Counting.............................................................17
6.3Precision and Reproducibility.......................................................................18

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6.4Benefits of True Volumetric Absolute Counting..........................................18
6.5Performing True Volumetric Absolute Counting.........................................19
7Optical Standard Setup – Parameters.................................................................21
7.12 Laser instrument .........................................................................................21
7.23 Laser instrument (high power laser 488 nm)............................................21
7.3Other parameter allocations possible ..........................................................22
8Optical Standard Setup – Flow geometry...........................................................23
9Instrument Settings ..............................................................................................24
9.1The Parameter Setup Dialog Box..................................................................24
9.1.1Trigger..............................................................................................................25
9.1.2Gain..................................................................................................................26
9.1.3Sample Speed..................................................................................................27
9.1.4Threshold: Lower Level (L-L)............................................................................29
10Disposal.................................................................................................................30
10.1Disposal of device..........................................................................................30
10.2Disposal of components................................................................................30
10.3Waste disposal ...............................................................................................30
10.4Sheath Fluid disposal ....................................................................................30
11Appendix................................................................................................................32
11.1Installation and uninstallation.......................................................................32
11.2Installation Requirements..............................................................................32
11.3Instrument Setup – Step by Step ..................................................................34
11.4The Flow Cuvette............................................................................................37
11.5Maintenance and Service...............................................................................39
11.5.1Maintenance..................................................................................................39
11.5.2Service ..........................................................................................................39
11.6Transport and Storage...................................................................................39
11.7Disposal...........................................................................................................39
11.8Laser Safety ....................................................................................................40
12Technical data.......................................................................................................41
12.1System.............................................................................................................41
12.2Optics ..............................................................................................................42
12.3Fluidics............................................................................................................43
12.4Electronics and Computer.............................................................................43
12.5Software ..........................................................................................................44

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Table of figures
Figure 1:Cells are separated in a suspension and stained with fluorescent markers.....9
Figure 2:Light from the laser(s) excites cell-bound fluorophores and is scatted by the
cells..................................................................................................................9
Figure 3:Fluorescence signals are displayed and analysed in histogram and/or dot plot
diagrams.........................................................................................................10
Figure 4:True Volumetric Absolute Counting by the Electrode Principle......................18
Figure 5:Exemplary demonstration of the optical bench of a CyFlow™ Space with 2
lasers and 8 parameters (left) and 3 lasers and 8 parameters (right). Other
laser light sources and parameter allocations are possible. D = detector (photo
multiplier)........................................................................................................22
Figure 6:Optical geometry at the laser interrogation (schematics)...............................23
Figure 7:Multiparameter instrument settings box .........................................................24
Figure 8:The Parameter Setup Dialog Box...................................................................24
Figure 9:Sample Transporting System of the CyFlow™ Space. ..................................28
Figure 10:Recommended placement of CyFlow™ Space components.........................32
Figure 11:Instrument Setup and periphery (CyFlow™ Space shown from the rear)......33
Figure 12:Computer connection on the CyFlow™ Space rear.......................................34
Figure 13:Power connection and light source switches on the left side of the CyFlow™
Space.............................................................................................................35
Figure 14:Tube connections for sheath and waste containers and cable connections for
the fluid level sensors on the right side of the CyFlow™ Space.....................36
Figure 15:Flow cuvette with sample, sheath fluid and waste connections. ....................37
Figure 16:Warning laser radiation ..................................................................................40
Figure 17:Additional explanation....................................................................................40

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1 Identification
1.1 Product information
Name CyFlow™ Space
Software FloMax®2.1x
REF CY-S-3001R
1.2 Manufacturer
Sysmex Partec GmbH
Arndtstraße 11 a-b,
02826 Görlitz,
Germany
Phone [phone]
Fax [phone]
[email protected]
www.sysmex-partec.com

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2 Introduction
2.1 What is the CyFlow™ Space?
The CyFlow™ Space is a fully equipped desktop Flow Cytometer (FCM). It features a
modular optical concept which allows using up to four different laser light sources and
between one and sixteen optical channels (parameters). The CyFlow™ Space allows easy
adaption of the optics to any application by simple exchange of optical filters and mirrors.
The CyFlow™ Space runs with a standard desktop PC. Data acquisition, instrument control,
and data analysis are controlled and performed by the FloMax®software.
2.2 What are the applications for which the CyFlow™ Space can be
used?
Together with the software, the CyFlow™ Space offers automation for routine use and
flexibility for research use for practically any flow cytometric application. The applications
cover e.g.:
Research based Immunophenotyping and other Blood Cell Analysis
Cell Counting / Rare Event Analysis
Cell cycle and cell proliferation Analysis
Detection and Analysis of Microorganisms and Viruses
Analysis of Environmental Samples
Fermentation Control
Particle Size and Fluorescence Distribution Analysis
2.3 What topics are covered by this manual?
The CyFlow™ Space Instrument Operating Manual covers the basic operation and
maintenance of the CyFlow™ Space instrument. This manual does not cover details related
to the software. Different software operating manuals are available.
2.4 What other manuals are available?
FloMax®– Acquisition and Instrument Control
FloMax®– Data Analysis: all aspects of on- and offline data analysis
CyPAD® with CyFlow™ Space Autoloading Station Operating Manual
Application Notes: for various applications
2.5 What should I know before operating the CyFlow™ Space?
This manual assumes you have basic knowledge about flow cytometry. In the best case a
flow cytometry expert is around - so let her/him help you. Basic books are available about
flow cytometry, which may help you as well (e.g. Howard M. Shapiro, Practical Flow
Cytometry. Wiley 2002).

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2.6 What standards were applied for CE compliance?
The system described in this manual is marked with a CE-mark, which
confirms the compliance with the essential requirements of the following
European Directives:
2014/35/EU on electrical equipment designed for use within certain
voltage limits
2014/30/EU on electromagnetic compatibility
2011/65/EU on the restriction of the use of certain hazardous
substances in electrical and electronic equipment
2.7 What typographical conventions are used in this manual?
Read the Operating Manual as well as the associated labelling thoroughly before using
the device. Please keep the Operating Manual for future reference.
The Operating Manual will help operating the device. Emphasis lays on installation, start-
up and safe operation of the device.
If you have any questions about the content of the Operating Manual or operating the
device, please contact your local Sysmex representative.
2.7.1 Symbols in this document
To make warnings more accessible, the following symbols are used to reference the kind
of hazard1.
Hazard
symbol Meaning
General hazards
Biological hazards
Laser radiation hazards
Electrical hazards
Beware of electrical currents and electrical shocks.
2.7.2 Safety information
Safety information and warnings are marked by symbols and signal words. Hazards are
categorized into four levels. All warnings are structured according to the same model. The
type of hazard is stated after the signal word. Then the consequences that may result
1These symbols are in black and white throughout this document, even though they appear in
colour on the device. Their meaning is unchanged.

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from the hazardous situation are described. And finally, how the hazardous situation can
be avoided. If applicable a hazard symbol is portrayed, to indicate the kind of hazard.
DANGER
Names the source of the hazard
Describes the consequences of the hazard. This will result in death or
severe personal injury.
Specifies the steps that must be taken to eliminate the hazard.
WARNING
Names the source of the hazard
Describes the consequences of the hazard. This could result in death or
severe personal injury.
Specifies the steps that must be taken to eliminate the hazard.
CAUTION
Names the source of the hazard
Describes the consequences of the hazard. This could result in minor
injury.
Specifies the steps that must be taken to eliminate the hazard.
NOTICE
Names the source of the hazard
Describes the consequences of the hazard. This could result in material
damage.
Specifies the steps that must be taken to eliminate the hazard.
INFORMATION
Gives further information about the device or procedure.

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3 Typical Steps of Particle Analysis
3.1 Preparation and Staining
For flow cytometric analysis, the cells (or other particles) must be separated from each
other in an aqueous suspension. Typically, if not generating sufficient optical signals by
themselves, the cells are then labelled by staining with one or more fluorescent dyes. The
fluorescent molecules bind to the cell substance of interest, e.g. CD4- and CD45-antibodies
bind cell surface antigens of leukocytes. The quantity of labelling cells with a specific dye is
proportional to the number of antigen molecules of a single cell. A good preparation with
suited reagents is the requirement for a precise analysis. A check of staining in a
fluorescence microscope can un-reveal preparation problems.
Figure 1: Cells are separated in a suspension and stained with fluorescent markers
3.2 Flow Cytometry Analysis
While passing through a flow cuvette one-by-one, the cells are individually illuminated by
the light spot of the laser. Due to the excitation, the dye molecules emit fluorescence of
characteristic colour (emission wavelength spectrum). This fluorescence light is separated
into colour ranges by means of optical filters. The intensity of each colour range is analysed
for each single cell.
Besides fluorescence, the intensity of light scattered by each cell can be measured. Scatter
light is measured in forward direction from the light source (forward scatter, FSC) and
sideward direction (side scatter, SSC). The scatter intensity is a measure of cell size and
morphology. Scatter light can be used to identify a cell before analysing its fluorescence,
but a cell can also be identified by fluorescence before analysing its scatter properties.
Figure 2: Light from the laser(s) excites cell-bound fluorophores and is scatted by the cells
3.3 Real-time Data Processing and Results
The light intensity of each parameter, and with it the quantity of substance of interest, is
assigned to one of up to 216 or 65536 quantity classes (channels). The classification is
performed in real-time, while the cells are passing the flow cuvette. Single parameter

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histograms show the number of cells within the channels. Two parameter dot plots show
the correlation between two cell properties.
Figure 3: Fluorescence signals are displayed and analysed in histogram and/or dot plot diagrams
3.4 Absolute Cell Counting
Since the CyFlow™ Space analyses all cells passing through the flow cell while precisely
monitoring the fluid volume of the sample, it allows volumetric counting during the analysis,
i.e. the determination of concentration of any cell subpopulation. Cell subpopulations can
even be defined and their concentration analysed at a later date, after reloading data from
a file.

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4 Safety
4.1 Laser radiation hazards
The device is a class 1 laser product, while the housing is unopened,
according to EN 60825-1. If the housing is opened and laser light is
emitted, the device is a class 3b laser product. Laser light exposure
causes damage to skin and eyes. Avoid direct exposure.
4.2 Electrical hazards
Please place the device in such a way, that the wiring is easily accessible
to unplug and does not suffer damages in any way. Only replace
removable wiring with adequate spare parts.
4.3 Alterations to the device
Unauthorized alterations to the device or the software result in risks
towards all user groups and the device itself. Unless permitted by the
manufacturer, the device must not be altered.

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5 Operating Basics
5.1 Switching on the CyFlow™Space
5.1.1 Check SHEATH and WASTE bottle
Make sure SHEATH bottle is filled with no more than 1600 ml of Sheath
Fluid and is closed tightly with the screw top. Tilt sheath bottle in order
to release air bubbles trapped in the yellow inline filter!
Make sure the WASTE bottle is empty.
It is recommended to replace the sheath fluid at least once a week or before any daily use.
5.1.2 Switch on the instrument
The CyFlow™ Space Flow Cytometer is operated with 100- 240 V AC.
Switch on main power at the left side of the instrument. The 488 nm
laser has its own power button close by. Also switch on the 488 nm
laser, please see chapter 10.2 Instrument Setup – Step by Step, Figure:
13.
638/640 nm lasers are software controlled or are switched by additional
labelled hardware switches next to the 488 nm laser power switch, e.g.
the 405 nm, the 375 nm and the 532 nm lasers.
5.1.3 Control of the 488 nm laser output power (for 200 mW high power laser only,
model Coherent Sapphire)
The 488 nm 200 mW high power laser can be adjusted in its operation
power between 50 and 200 mW. For power selection please double click
the desktop icon shown on the left (488-200).
To change selected laser power please type in:
p=value enter
To inquire about actual selected laser power type in:
?p enter
5.1.4 Switch on peripheral devices
Switch on printer and/or sorter module.
5.1.5 Switch on computer
Finally, switch on computer by the Computer Power switch.

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The display shows the Windows®desktop.
5.1.6 Start instrument operating software
Double click FloMax®icon to start FloMax®CyFlow™ Space operation software.
After a few seconds, the display shows the FloMax®Welcome window.
By confirming with “OK” the CyFlow™ Space is initialized and the last used instrument
settings are loaded.
The display shows empty histograms.
5.1.7 Priming the instrument (initial cleaning)
Use speed 5. Run a tube filled with 1.6 ml Cleaning Solution (REF 04-4009_R) until the
system stops. Remove sample tube and run a second tube with Sheath Fluid (04-4007_R)
for 2 minutes.
The CyFlow™ Space is ready for acquisition.
DANGER
The waste may contain biohazardous material (infectious cells, dyes). To
minimize biohazards, fill about 25 ml of Hypochlorite Solution (REF 04-
4012_R) into the waste container. When emptying the container, be sure
not to come in contact with the fluid. In case of accidental contact, wash
your skin throughout with soap and disinfectant.
5.2 Multi laser Measurements
NOTICE
Alignment of lasers must only be done by Sysmex representatives or by
authorized service personnel.
The CyFlow™ Space Flow Cytometer can be equipped with one up to 4 laser light sources.
Actual optical setup of your instrument may therefore vary. For questions concerning the
laser setup of this instrument, please contact your local Sysmex representative.
In order to minimize crosstalk between multiple light sources, laser beams from different
lasers are focused on different positions (spots) of the flow cuvette. The CyFlow™ Space
Flow Cytometer supports either 2 laser spots (when employing FloMax®versions 2.x) or 3
laser spots (when employing FloMax®version 3.x). The blue 488 nm laser as leading laser
of the CyFlow™ Space is always located on spot 1. Additional lasers are located on spot 2
and spot 3, respectively. Instruments without 488 nm laser, e.g. only with UV-LED or with
532 nm laser will use that light source on spot 1.

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All installed light sources except 638 nm or 640 nm diode lasers are
switched by hardware switches located at the left side of the flow
cytometer. The laser switches are located above the instruments main
on/off switch. Lasers will be only on power when the main instrument
power is on. Red laser diodes, 638-640 nm are switched by a software
switch in FloMax®.
Select “Acquisition” from the upper command line and click on Laser No.
2 to power the laser on. The tick indicates that the laser is switched on.
The software switch Laser No. 3 has no function in CyFlow™ Space
systems.
Laser operation status is saved in Instrument settings files.
In the standard configuration all particles pass through laser
spot no. 1 of the 488nm laser – the leading laser – first and
then through the laser(s) at spot no. 2 and no. 3 (if present).
Therefore, all parameters derived from a laser located at a spot
other than no. 1 require a time delay. Time delay of detectors
is set in the Parameter Setup dialog box (see chapter 9.1).
5.3 Starting a Measurement
NOTICE
Please see “FloMax®- Acquisition and Instrument control” on how to
change instrument settings.
Make sure your flow cytometer is ready for analysis and the operating
software is prepared for the measurement.
1. Prepare sample according to the Application Notes and
preparation procedures, resp. Use not less than 830 µl for true
volumetric absolute counting and not more than 2.8 ml. For smaller
amounts of sample use Small Volume Sample Tubes (REF 04-2010, however these
tubes are not compatible with the True Volumetric Absolute Counting option).
2. Insert sample tube into the sample port until you recognize a
"click". The sample should be fully mounted within a second.

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Now the measurement (acquisition) starts automatically – the operating software indicates
Prerun, Stabilize, Run and Count phases. Instrument status is also highlighted by LEDs
on the front side of the CyFlow™ Space.
In the Prerun phase, cells are quickly transported into the flow cuvette.
During the Stabilize phase, sample flow stabilizes and the flow reaches the final set
sample injection rate.
In the Run phase, cells are analysed and data are acquired.
In the Count phase, cells are counted for a given volume. Do not change instrument
settings in the count phase! Do not clear data!
After the count phase, the acquisition finishes automatically. To finish the
acquisition before, click the END button, or simply remove the sample tube
from the sample port. In any case the instrument initiates now the
REPOSITION phase to prepare the instrument for the next analysing cycle.
or
A Biosafety cleaning cycle is started.
The system status changes to Clean and then Ready.
The system is ready for the next sample.
NOTICE
When removing a tube from the sample port, the BioSafety Cleaning
system is activated to prevent dropping and contamination by hazardous
material and to clean the sample tube. During Cleaning and Reposition
phases FloMax®does not accept any actions. Instrument operation can
only be continued after finalization of both phases.
To restart an acquisition without removing the sample tube, click onto start.
or
3. To save results, click on the save file icon.

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5.4 Switching off the CyFlow™ Space
NOTICE
Final cleaning with sheath fluid is an obligation to avoid clogging of the
flow cuvette.
Make sure data from the last acquisition have been saved.
1. Clean the flow cuvette and connecting tubes
Use speed 5.
a) Daily cleaning procedure:
Run 1.6 ml of Cleaning Solution (REF 04-4009_R) until sample tube is empty. Run 1.6 ml
of Decontamination Solution (REF 04-4010_R) for one minute. Press CLEAN button once
to start manually a cleaning cycle. Run 1.6 ml Sheath Fluid (REF 04-4007_R) for two
minutes. Press STOP.
b) Weekly cleaning procedure:
Run 1.6 ml of Decontamination Solution. Stop the instrument by pressing END. Incubate
for 15 minutes. Re-start the system by pressing START and let it run to the end. Run the
system with 1.6 ml Cleaning Solution until the sample tube is empty. Run 1.6 ml Sheath
Fluid for two minutes. Press STOP.
2. Protect the sample port
Leave a test tube filled with sheath fluid connected to the sample port – this avoids drying
and crystallization of any remaining material in the tubing.
3. Quit the operating software
Click onto the close button [X] in the upper right corner of the window (or select File...Exit
in the menu).
4. Shut down the computer
Click with the left mouse button onto the Windows®"Start" button in the taskbar - the
Windows menu is displayed.
Click "Turn Off Computer..." - the shutdown window appears.
Click "Turn Off" in the Shutdown Window.
5. Switch off printer/sorter module
6. Switch off instrument
Switch off main power and the individual lasers at the left side of the instrument.

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6 True Volumetric Absolute Counting – Overview
Analysis of concentrations of cells and particles, loosely called Absolute Counting, can be
of significant interest for various studies e.g. monitoring the number of cells in cell cultures
or biotechnological processes.
6.1 Non-volumetric Flow Cytometric Techniques for Absolute
Counting
i) Dual Platform Technique
To determine the concentration of a sample flow cytometers (FCM) are frequently used in
combination with Haematology Counters.
Disadvantages:
Two instruments required
This technique relies on the assumption that the number of cells seen by the FCM and
counted by the Haematology Counter are identical.
ii) Single Platform Technique with Reference Beads
An alternative method employs reference beads of known concentration, which are added
to the sample of unknown concentration.
Disadvantages:
Depends on the accuracy of the specified beads concentration
Constant running costs for the reference beads
6.2 True Volumetric Absolute Counting
To overcome the drawbacks of the Dual- and Single-Platform Technique with reference
beads, Sysmex Partec instruments additionally offer an alternative way of absolute cell
counting, which is based directly on the basic definition of a concentration by precisely
counting the number Nof cells suspended in a purely mechanically defined volume V.
𝑐
𝑁
𝑉
i) Precise Counting: Determination of N
For True Volumetric Counting the precise detection of cells is an essential. This requires
fast recognition and analysis of the events by electronics and computer. All Sysmex Partec
instruments are specifically designed to minimize counting losses by providing direct
connection between computer and electronics, which avoids dead-times involved in
traditional FCM designs and instrument interfaces. This reduces the probability of a
counting loss for typical event rates below 2%.

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ii) Electrode-Principle: Determination of V
The method for True Volumetric Absolute Counting supported by the CyFlow™ Space is
based on the precise measurement of a fixed sample volume by means of two electrodes.
During analysis by the CyFlow™ Space, the sample liquid loses contact first from the upper
and then from the lower electrode. These events trigger a START and a STOP signal,
respectively which is stored by software.
Figure 4: True Volumetric Absolute Counting by the Electrode Principle
The volume Vof sample liquid analysed between the START and STOP signal is physically
defined by the distance of the two electrodes and the diameter of the sample tube. The
analysed volume is exactly 200 µl (= counting volume).
Effects of the sample meniscus in the tube are eliminated because the START and STOP
electrodes are arranged symmetrically. The sample conductivity does not influence the
volumetric measurement, as long as the fluids can be detected electrically.
6.3 Precision and Reproducibility
Precision and reproducibility of the Sysmex Partec instruments can be demonstrated and
checked with Count Check beads (low, medium, high) (REF 05-4010).
6.4 Benefits of True Volumetric Absolute Counting
Sysmex Partec instruments can perform traditional Dual- and Single-Platform counting and
unique True Volumetric Absolute Counting, the latter offering unique benefits:
i) Precision
A high precision of better than 5% is guaranteed by precise counting and mechanical
volume measurement. The counting reproducibility is better than 2% rel. standard deviation.

CyFlow™Space | Operating Manual | March 2021 19
ii) No Errors Due to Calibration
Instability of beads suspensions over time or counting statistics of beads cannot influence
the counting results, since no beads are used.
iii) Less Preparation or Setup Time
The sample concentration is directly analysed by the FCM without any instrument
calibration or additional sample preparation step.
iv) Less Analysis Time
Concentration results for subpopulations, as defined by gates, are immediately displayed
on the instrument screen. No additional analysis steps, e.g. setting gates for beads, are
required.
v) Less Expenses
No reference beads required.
6.5 Performing True Volumetric Absolute Counting
Sedimentation and Count Time
Cells or other particles may tend to sediment inside the sample tube depending on their
density in relation to the surrounding suspension solution. Significant sedimentation can
take place within minutes. The CyFlow™ Space analyses the concentration at the location
of the sample uptake, where it can vary over time due to sedimentation or de-sedimentation
effects. Consequently, avoid too long count phases due to low sample speed
(e.g. below 2 µl/s). Count Time typically should not exceed 2-4 minutes.
1. Shake sample for re-suspension of particles, but avoid producing air bubbles on the
surface of the liquid.
2. Fill 850 µl of sample into a test tube (instrument setup should be set to: pre-run 2
sec, stabilize 2 sec).
3. Insert sample tube onto sample port.
The acquisition starts and performs:
Pre-run
stabilize
run
count: particles are counted during the count phase.
The acquisition stops automatically. Biosafety cleaning is done and Ready is
displayed when the system is ready to analyse the next sample.
4. Define subpopulations by regions and gates to analyse concentrations
(concentration will be displayed for each subpopulation).
During the count phase, user interference has to be avoided!

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NOTICE
If in Acquisition Setup/Acquisition Control, the function Clear Histograms
on Count Start is enabled all data acquired during RUN phase prior to the
COUNT phase will be deleted. The final fcs data file will contain only the
data acquired during COUNT phase of 200 µl sample. Analysing the data
with 3rd-party fcs software will allow the calculation of concentrations. To
achieve numbers per ml multiply the count with the factor 5 (count in 0.2
ml x 5 = count / ml).
If the function, Clear Histograms on Count Start is disabled FloMax®will
label the data set acquired during COUNT phase but will maintain all data
of RUN and COUNT phase. In FloMax®Cell concentration will be
calculated on basis of the data label. In 3rd party software a calculation of
concentration is not possible as a read out of the data label is not
possible.
CAUTION
When analysing data without TVAC make sure Clear Histograms on
Count Start is disabled. Otherwise data are lost when entering COUNT
phase!