SYSMEX CyFlow Cube 6 V2m User manual

With CyView™ 1.8
Doc. No.: CY-S-3061R-V2mIFUEN | Rev: 002 | Rev. date: 03-02-2020 | EN | CN 1432
CyFlow™ Cube 6 V2m
Operating Manual

Table of contents
CyFlow™Cube 6 V2m | Operating Manual | February 2020
3
Table of contents
1Introduction.......................................................................................................8
1.1 About this Operating Manual...............................................................................8
1.2 Typographical conventions .................................................................................8
1.2.1 Safety information...............................................................................................8
1.2.2 Text highlights.....................................................................................................9
1.3 General conditions............................................................................................10
1.4 CE compliance..................................................................................................10
1.5 Sysmex representatives....................................................................................11
1.6 Authorised distributors......................................................................................11
2Safety...............................................................................................................12
2.1 Intended Use ....................................................................................................12
2.2 Prohibited use...................................................................................................12
2.3 Reasonably foreseeable misuse.......................................................................12
2.4 User definition...................................................................................................12
2.4.1 User qualification groups...................................................................................12
2.4.2 User Qualification Matrix...................................................................................13
2.5 Environmental conditions..................................................................................13
2.6 Prohibition and warning signs on the device .....................................................13
2.7 Personal protective equipment..........................................................................14
2.8 General safety ..................................................................................................14
2.9 Laser radiation hazards ....................................................................................14
2.10 Electrical hazards..............................................................................................15
2.11 Alterations to the device....................................................................................15
3Overview..........................................................................................................16
3.1 Device...............................................................................................................16
3.2 Components of the device ................................................................................18
3.2.1 Control elements...............................................................................................18
3.2.2 Computer interface ports...................................................................................18
3.2.3 Sheath Fluid and Waste bottle..........................................................................18
3.2.4 Sample port with electrodes..............................................................................19
3.2.5 Type plate.........................................................................................................19
3.3 Scope of delivery ..............................................................................................20
3.4 Accessories and spare parts.............................................................................21
4Transport and storage....................................................................................22
5Installation.......................................................................................................23
5.1 Positioning the device.......................................................................................23
5.2 Installation and uninstallation............................................................................23
6Operation.........................................................................................................24
6.1 Start-up procedure............................................................................................24
6.1.1 Switching on the device....................................................................................24
6.1.2 Logging in.........................................................................................................24

Table of contents
4
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
6.1.3 Priming the device ............................................................................................25
6.1.4 Performing a quality check................................................................................26
6.1.5 Analysing a quality check..................................................................................27
6.2 Performing a measurement (Sample Port)........................................................28
6.3 Saving acquired data........................................................................................29
6.4 Performing intermediate cleaning......................................................................29
6.5 Shutdown procedure.........................................................................................30
6.5.1 Shutdown cleaning............................................................................................30
6.5.2 Switching off the device ....................................................................................31
6.6 Software operation............................................................................................31
7Troubleshooting..............................................................................................32
7.1 Fault, cause and remedy...................................................................................32
7.1.1 Sheath Fluid, waste and other fluids .................................................................32
7.1.2 Calibration and Count Check Beads (low, medium, high)..................................32
7.1.3 Measurements and data acquisition..................................................................33
8Maintenance....................................................................................................34
8.1 Maintenance overview......................................................................................34
8.2 Maintenance procedures...................................................................................35
8.2.1 Sheath Fluid refilling .........................................................................................35
8.2.2 Inline Filter renewal...........................................................................................35
8.3 Other maintenance ...........................................................................................36
8.3.1 Device cleaning ................................................................................................36
8.4 Technical service..............................................................................................37
8.4.1 Preventive maintenance....................................................................................37
8.4.2 Defective device ...............................................................................................37
9Disposal...........................................................................................................38
9.1 Disposal of device.............................................................................................38
9.2 Disposal of components....................................................................................38
9.3 Waste disposal .................................................................................................38
9.4 Sheath Fluid disposal........................................................................................38
10 Technical data.................................................................................................40
10.1 Environmental conditions..................................................................................40
10.2 Device...............................................................................................................40
10.3 Optics ...............................................................................................................41
10.4 Fluidics .............................................................................................................42
11 Software ..........................................................................................................43
11.1 User Management ............................................................................................43
11.1.1 Login..........................................................................................................43
11.1.2 User Administration....................................................................................43
11.2 Overview...........................................................................................................45
11.2.1 Title bar......................................................................................................45
11.2.2 Drop down menu .......................................................................................45
11.2.3 Main window..............................................................................................46

Table of contents
CyFlow™Cube 6 V2m | Operating Manual | February 2020
5
11.2.4 Controls.....................................................................................................47
11.3 Data management ............................................................................................48
11.3.1 Open (.fcs).................................................................................................48
11.3.2 Save (.fcs) .................................................................................................48
11.3.3 Open Configuration....................................................................................48
11.3.4 Save Configuration ....................................................................................48
11.4 Data acquisition controls...................................................................................49
11.4.1 Acquisition controls....................................................................................49
11.4.2 Device real time display.............................................................................49
11.5 Settings.............................................................................................................50
11.5.1 Compensation............................................................................................51
11.5.2 Plots ..........................................................................................................52
11.5.3 Results.......................................................................................................53
11.5.4 Regions .....................................................................................................54
11.5.5 Acquisition.................................................................................................59
11.5.6 Measure (Sample Port)..............................................................................60
12 Annex...............................................................................................................63
12.1 Abbreviations....................................................................................................63
12.2 List of software commands ...............................................................................64

Table of figures - Figures
6
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
Table of figures - Figures
Figure 1: Front and left side of the device .......................................................................16
Figure 2: Ports for external devices.................................................................................17
Figure 3: Rear side of the device.....................................................................................17
Figure 4: Sample port at the right side of the device .......................................................18
Figure 5: Type plate........................................................................................................19
Figure 6: Selecting Prime Mode......................................................................................25
Figure 7: Selecting QC Mode..........................................................................................26
Figure 8: Selecting Work Mode.......................................................................................29
Figure 9: Selecting FCS-File Storage..............................................................................29
Figure 10: Intermediate Cleaning....................................................................................30
Figure 11: Selecting Shutdown Cleaning Mode...............................................................30
Figure 12: Login and system check window....................................................................43
Figure 13: User administration window (exemplary)........................................................44
Figure 14: Main window (5Pl.cvc85 configuration active)................................................46
Figure 15: File operations ...............................................................................................47
Figure 16: Device real time display .................................................................................47
Figure 17: Acquisition controls........................................................................................47
Figure 18: Intermediate Cleaning....................................................................................47
Figure 19: Compensation tab..........................................................................................51
Figure 20: Plots register (Histogram)...............................................................................52
Figure 21: Plots register (Dot plot)...................................................................................52
Figure 22: Results register..............................................................................................53
Figure 23: Creating a quadrant region.............................................................................55
Figure 24: Creating a vertical Region..............................................................................55
Figure 25: Create a polygonal region..............................................................................56
Figure 26: display options...............................................................................................57
Figure 27: Show Regions (density colored).....................................................................57
Figure 28: Show Regions (unicolored)............................................................................57
Figure 29: Show all events + Regions (unicolored) .........................................................57
Figure 30: Region properties...........................................................................................58
Figure 31: Acquisition register.........................................................................................59
Figure 32: Measure register............................................................................................60
Figure 33:Acquisition process for all Sample Port Measure Modes.................................61
Figure 34: Workstep register...........................................................................................62

Table of figures - Tables
CyFlow™Cube 6 V2m | Operating Manual | February 2020
7
Table of figures - Tables
Table 1: Text highlights.....................................................................................................9
Table 2: Representative's contacts .................................................................................11
Table 3: User Qualification Matrix ...................................................................................13
Table 4: Safety signs on the device.................................................................................14
Table 5: Personal protective equipment (PPE)................................................................14
Table 6: Signs on the type plate of the device.................................................................19
Table 7: Scope of delivery and operation........................................................................20
Table 8: Reagents...........................................................................................................21
Table 9: Accessories and spare parts.............................................................................21
Table 10: Measure Modes ..............................................................................................28
Table 11: Maintenance activities.....................................................................................34
Table 12: Roles and privileges........................................................................................44
Table 13: Drop down menu structure for each configuration (Mainuser) .........................45
Table 14: Drop down menu structure for each configuration (User).................................46
Table 15: Acquisition controls .........................................................................................49
Table 16: Device real time display elements...................................................................49
Table 17: Manual Sample Port Measure Modes..............................................................62

1 Introduction
8
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
1 Introduction
1.1 About this Operating Manual
This Operating Manual will help you to operate your CyFlow™ Cube device. Emphasis is
placed on the installation, start-up and safe operation of the device.
The basic principles of flow cytometry can be found in the scientific literature (e.g. Howard
M. Shapiro, Practical Flow Cytometry, Wiley 2003).
This Operating Manual comes with the device. Read this Operating Manual thoroughly
before using the device. Please keep this Operating Manual for future reference.
This Operating Manual is divided in two parts. The first part, chapter 1 to 10, gives the
user all necessary information for safe handling, scope of delivery, to set up the device
and how to operate the device properly. The general use of the software and the typical
workflows are described in chapter 6, this chapter also is suitable to get to know the
functioning of your device.
In the second part, chapter 11, you will find a description of all software functions and a
detailed description of the user interface. This chapter is for reference.
There are product data sheets and assay manuals for the reagent kits and general
application notes available. Please also refer to these documents.
If you have any questions about the content of this manual or the use of the device,
contact your local Sysmex representative.
1.2 Typographical conventions
1.2.1 Safety information
Safety information and warnings are marked by symbols and signal words. Hazards are
categorised 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
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.

1 Introduction
CyFlow™Cube 6 V2m | Operating Manual | February 2020
9
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.
1.2.2 Text highlights
To ease the reading flow, the following kinds of text highlights are used in this manual.
Table 1: Text highlights
Style
Meaning
[Brackets]
Describes an interactive element of
software. This includes paths, buttons and
tabs.
Example: Press the [Start] button.
Italic 11 pt
References instances for interaction
(names thereof) in instructions.
Example: Push the On switch on the top of
the device.
It may also be used as chapter reference.
Example: For more information, please
refer to chapter 11.1 User Management.
Italic 8 pt
Used for captions of figures (figures,
tables).
Example: Figure 20: General overview, Table 2:
Safety information
It may also be used for additional
descriptive text.
Example: Electrical hazards Beware of
electrical currents and electrical shocks
“Quotes”
Describes reference sources.
Example: Please consider our “General

1 Introduction
10
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
Terms and Conditions of Sale and
Delivery” […]
Bold
Describes headlines of subtopics, which
need no independent chapter.
Example: Qualified personnel
Describes also specially highlighted
keywords.
Example: Names the source of the
hazard
Underlined
References external hyperlinks.
Example: www.sysmex-partec.com
1.3 General conditions
Ensure that the following conditions are met when working with the device:
•This Operating Manual must be read prior to using the device.
•Report any recurring faults or problems to your local Sysmex representative.
Sysmex Partec GmbH does not accept liability for any damage or personal injury arising
from:
•Unauthorised modifications or manipulation of the device;
•Improper use;
•Use of non-approved accessories;
•Improper handling;
•Non-compliance with this Operating Manual.
Not following these conditions will impair the safe use of the device, its components and
the safety of the user.
Sysmex Partec GmbH regulates the intended purpose of its products in the instructions
for use. The manufacturer assumes no responsibility for any application deviating from
the instructions for use stated in the product documentation (Off Label Use). We
expressly emphasize that in the event of use contrary to the intended purpose the liability
rests solely with the user.
1.4 CE compliance
The device is marked with a CE mark of conformity, which confirms the
compliance with the essential requirements of the following European
Directives:
•2014/35/EC on electrical equipment designed for use within certain
voltage limits;
•2014/30/EC on electromagnetic compatibility

1 Introduction
CyFlow™Cube 6 V2m | Operating Manual | February 2020
11
1.5 Sysmex representatives
Find address and contact data of your local Sysmex representative also at
www.sysmex-partec.com.
Table 2: Representative's contacts
Europe, Middle East and Africa
Sysmex Europe GmbH
Bornbarch 1,
22848 Norderstedt,
Germany
Phone: [phone]
Fax: [phone]
Americas
Sysmex America, Inc.
577 Aptakisic Road,
Lincolnshire,
IL 60069,
U.S.A.
Phone: [phone]
Fax: [phone]
Asia Pacific
Sysmex Asia Pacific Pte Ltd.
9 Tampines Grande #06-18,
Singapore 528735
Phone: [phone]
Fax: [phone]
China
Sysmex Shanghai Ltd.
9th Floor, AZIA Center 1233, Lijuazui Ring Road,
Shanghai 200120, PRC
Phone: [phone]
Fax: [phone]
Japan
Sysmex Corporation
1-5-1 Wakinohama-kaigandori, Chuo-ku, Kobe, Hyogo
651-0073, Japan
Phone: [phone]
1.6 Authorised distributors
To receive an overview of authorised distributors nearby, please contact your local
Sysmex representative.

2 Safety
12
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
2 Safety
2.1 Intended Use
The system is intended for the industrial market. Therefore, the CyFlow™Cube 6 V2m is
a device, which is not for use in diagnostic procedures. The CyFlow™Cube 6 V2m
provides the basis for the following applications:
•Microbiological analysis;
•Fermentation control;
•Bead-based assays for research purposes;
•Particle and cell concentration analysis based on True Volumetric Absolute
Counting;
•Particle size and fluorescence distribution analysis.
2.2 Prohibited use
Never use the system with organic solvents as this can destroy the fluidic system. Only
use particles with a diameter < 100 μm. It is recommended to use Sysmex Partec
accessories and consumables only.
Never for use in diagnostic procedures. Not for use with biohazardous substances.
2.3 Reasonably foreseeable misuse
Only use this device as a sensitive measuring device.
•Do not use as personal computer.
•Do not use with chemicals harmful to the device (household sanitisers etc.).
•Do not place any objects on top of the CyFlow™ Cube 6 V2m.
2.4 User definition
The device may only be used by personnel properly instructed in its handling. The
manufacturer, authorised distributor or a person authorised by the operator (and trained
by either the manufacturer or an authorised distributor) must provide instruction in
handling the device with the help of this Operating Manual.
The device is aimed for a large spectrum of users, from beginners to experts. In the best
case an expert is around –so, if you are a beginner, let her/him help you.
Access to the device is not permitted to insufficiently qualified personnel.
2.4.1 User qualification groups
Instructed personnel
Instructed personnel have to be trained by qualified personnel or personnel authorised by
Sysmex to operate the device within the scope of the intended use. They have to be able
to assess if the device can be safely used and are proficient in handling hazardous
substances.

2 Safety
CyFlow™Cube 6 V2m | Operating Manual | February 2020
13
Qualified personnel
Qualified personnel have in-depth knowledge of the device and the applications. They
have been trained by personnel authorised by Sysmex to operate the device, to perform
cleaning and certain maintenance operations. Qualified personnel have a scientific or
technical education and several years of experience in flow cytometry and are proficient in
handling hazardous substances.
Authorised service personnel
Personnel conducting technical service must be authorised by Sysmex. Please contact
the service department of your local Sysmex representative.
2.4.2 User Qualification Matrix
The User Qualification Matrix represents an overview for activities associated with the
device during its life cycle.
Table 3: User Qualification Matrix
Activity
Instructed
personnel
Qualified
personnel
Authorised service
personnel
Transport
✓
✓
Installation
✓
Operation
✓
✓
✓
Cleaning
✓
✓
✓
Decontamination
✓
✓
Maintenance
✓
✓
✓
Technical
Service
✓
Uninstalling
✓
✓
Packing
✓
✓
Disposal
✓
✓
2.5 Environmental conditions
The device may only be used in laboratories. The specified ambient temperature and
relative humidity conditions must be maintained at all times.
The laboratory environment must be clean. Direct sun light should be avoided.
•Operating temperature range: 15 °C to 30 °C
•Relative humidity: between 20 % and 85 %, non-condensing.
2.6 Prohibition and warning signs on the device
The following prohibition and warning signs have been affixed to the device.

2 Safety
14
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
Table 4: Safety signs on the device
Symbol
Description
Laser radiation
General hazards
Additional laser labelling
2.7 Personal protective equipment
The following personal protective equipment is recommended to operate the device.
Table 5: Personal protective equipment (PPE)
Item
Description
Gloves must be worn at all times.
Lab coat must be worn at all times.
2.8 General safety
The system operates with the reagents stated in the scope of operation,
such as Hypochlorite Solution and Decontamination Solution. For further
information please see chapter 3.3 Scope of delivery.
The waste bottle collects all used substances, its contents are not able to
re-enter the fluidic system by design.
Important information regarding the safe handling, transport and disposal
of reagents is contained in their Safety Data Sheets.
Always meet the national and international guidelines and regulatory
standards for PPE (personal protective equipment).
Find Safety Data Sheets to our products at www.sysmex-partec.com
2.9 Laser radiation hazards
The device is equipped with a class IIIb laser unit. If the housing of the
device is closed, the laser class of the laser unit is classified as a laser
class I (EN 60825-1).

2 Safety
CyFlow™Cube 6 V2m | Operating Manual | February 2020
15
Only authorised service personnel is allowed to open the housing of the
device.
Laser light of laser class IIIb can be emitted if the housing of the device is
damaged or the laser protection is removed. Serious injuries of the eyes
and the skin are the results of emission by laser light of class IIIb lasers.
2.10 Electrical hazards
The main electrical source used must be grounded or earthed due to local
specifications. The mains cable carries out the protective earthing. Please
place the device in such a way that the electrical power cord can easily be
unplugged. Removable power cords should not be replaced by inadequate
wiring.
Beware of possible voltage when changing the fuse (T 3.15 A 250 V).
2.11 Alterations to the device
Unauthorised alterations to the device can result in risks and hazards. Therefore, unless
expressly permitted by the manufacturer, no alterations may be made to the device.
If the device is altered, appropriate tests and trials must be performed to ensure the
continued safe use of the device.

3 Overview
16
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
3 Overview
3.1 Device
NOTICE
Glass parts breakable
Objects put on the glass top or display of the device damage or may
break the glass.
•Do not place any objects on or scratch the glass top or display of the
CyFlow™ Cube.
Your CyFlow™ Cube device is a fully equipped desktop flow cytometer which features a
modular optical concept. This allows a 488 nm laser to be used as light source and the
detection of up to 5 optical channels (parameters).
The device allows the optics to be optimised for any application simply by exchanging
optical filters and mirrors. The device runs with an internal computer. Data acquisition,
device control, and data analysis are controlled and performed by the software.
Figure 1: Front and left side of the device
#
Item
#
Item
1
Sample port
3
On switch
2
Ports for external devices
4
Display

3 Overview
CyFlow™Cube 6 V2m | Operating Manual | February 2020
17
Figure 2: Ports for external devices
#
Item
#
Item
#
Item
1
USB port
2
HDMI port
3
Serial port
4
Ethernet port
5
USB port
6
Ethernet port
7
Audio port
8
Audio port
9
USB port
10
USB port
11
USB port
12
DVI port
13
Keyboard port
14
Mouse port
Figure 3: Rear side of the device
#
Item
#
Item
1
Tubing connection Waste bottle
5
Vacuum tubing connection Waste
bottle
2
Sensor cable connection Waste
bottle
6
Connector socket power cord
3
Sensor cable connection Sheath
Fluid bottle
7
Power switch
4
Tubing connection Sheath Fluid
bottle
8
Fuse insert (T 3.15 A 250 V)

3 Overview
18
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
Figure 4: Sample port at the right side of the device
#
Item
#
Item
1
Start electrode
4
Sample holder
2
Stop electrode
5
Sample port injection needle
3
Sample tube
3.2 Components of the device
The device contains the flow cuvette, the optics, the fluidics, the computer unit and data
acquisition electronics. No work steps inside the device are necessary.
The main components of the device are:
3.2.1 Control elements
The device is operated via keyboard and mouse. The On switch is located on the top,
when the display of the device is lifted up. The power switch is located on the rear side of
the device.
3.2.2 Computer interface ports
The computer interface ports connect the device with additional optional electronic
devices. They are located on the left side of the device.
3.2.3 Sheath Fluid and Waste bottle
The Sheath Fluid transports the sample flow during the measurement. The connecting
ports for the Tubing of the Sheath Fluid bottle and Waste bottle are located on the rear
side of the device. The Sheath Fluid bottle and Waste bottle are located next to the
device.

3 Overview
CyFlow™Cube 6 V2m | Operating Manual | February 2020
19
3.2.4 Sample port with electrodes
The Sample port is located on the right side of the device.
The Sample port holds the Sample tube during the measurement. The start and stop
electrodes of the Sample port allow the independent determination of exactly 200 μl
volume for each sample. The method of True Volumetric Absolute Counting (TVAC)
supported by CyFlow™ Cube systems is based on the precise measurement of a fixed
sample volume.
3.2.5 Type plate
The type plate is attached to the rear side of the device.
The content of the type plate varies according to the system configuration. The system
configuration is represented by an addition to the REF number (e.g. CY-S-3061R-V2m).
Figure 5: Type plate
Table 6: Signs on the type plate of the device.
#
Item
Manufacturer
accompanied by the name and the address of the manufacturer
Manufacturing Date
Serial Number
Reference Number
CE mark
Waste of Electrical and Electronic Equipment
Consult Instructions For Use

3 Overview
20
CyFlow™ Cube 6 V2m | Operating Manual | February 2020
#
Item
Caution
Keep Away From Sunlight
Temperature Limitation
upper and lower limits of temperature, here 15°C to 30°C
Humidity Limitation
upper and lower limits of humidity, here 20% - 85%
3.3 Scope of delivery
The following items are available through Sysmex and distributors authorised by Sysmex,
also the list should help the user identifiy what is already part of the scope of delivery.
Table 7: Scope of delivery and operation
Item
Scope of delivery
Scope of operation
CyFlow™ Cube 6 V2m
(device only)
✓
✓
Operating Manual
✓
✓
Power cord
✓
✓
Sheath Fluid bottle 1 l
✓
✓
Waste bottle 1 l
✓
✓
Cap for 1 l glass Sheath
Fluid bottle
✓
✓
Cap for 1 l glass Waste
bottle
✓
✓
Inline Filter
✓
✓
Sample tubes 3.5 ml
✓
FCS Express dongle
✓
✓
Keyboard
✓
✓
Mouse
✓
✓
Sheath Fluid with tap
✓
Cleaning Solution
✓
Decontamination Solution
✓
Hypochlorite Solution
✓