Related Manuals for SYSMEX CyFlow Space
Summary of Contents for SYSMEX CyFlow Space
- Page 1 ™ CyFlow Space Operating 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...
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Page 2: Table Of Contents
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 ... - Page 3 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 ...
- Page 4 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 ......................
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Page 5: Identification
1 Identification 1.1 Product information Name CyFlow™ Space ® Software FloMax 2.1x CY-S-3001R 1.2 Manufacturer Sysmex Partec GmbH Arndtstraße 11 a-b, 02826 Görlitz, Germany Phone +49 3581 8746-0 Fax +49 3581 8746-70 info@sysmex-partec.com www.sysmex-partec.com ™ CyFlow Space | Operating Manual | March 2021... -
Page 6: Introduction
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. -
Page 7: What Standards Were Applied For Ce Compliance
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... - Page 8 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.
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Page 9: Typical Steps Of Particle Analysis
3 Typical Steps of Particle Analysis 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. -
Page 10: Absolute Cell Counting
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 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. -
Page 11: Safety
4 Safety 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. -
Page 12: Operating Basics
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. -
Page 13: Start Instrument Operating Software
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. -
Page 14: Starting A Measurement
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. - Page 15 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. ...
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Page 16: Switching Off The Cyflow™ Space
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. -
Page 17: True Volumetric Absolute Counting - Overview
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. -
Page 18: Precision And Reproducibility
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). Benefits of True Volumetric Absolute Counting... -
Page 19: Performing True Volumetric Absolute Counting
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. - Page 20 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 3 -party fcs software will allow the calculation of concentrations.
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Page 21: Optical Standard Setup - Parameters
7 Optical Standard Setup – Parameters NOTICE If required by a specific application, the optical standard setup can be optimized by exchanging preassembled removable mirror/filter blocks. This is a matter of seconds and does not require readjustments. The CyFlow™ Space flow cytometer can be configured as flow cytometer or as a flow cytometer with cell sorting option. -
Page 22: Other Parameter Allocations Possible
FL5: red fluorescence II (APC) FL6: blue fluorescence (CFP, Alexa 405) 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. -
Page 23: Optical Standard Setup - Flow Geometry
8 Optical Standard Setup – Flow geometry waste side scatter fluorescence forward scatter laser beam (hydrodynamic focusing) sheath fluid sample Figure 6: Optical geometry at the laser interrogation (schematics) Laser light is focused into the flow cell. Laser light scattered from the particles is detected in the forward direction range (forward scatter). -
Page 24: Instrument Settings
9 Instrument Settings Instrument settings are used to optimize the CyFlow™ Space acquisition for the particles of interest. The adjustments cover the gains of the optical detectors, e.g. the photomultiplier high voltages, the amplification mode (lin, 3 or 4 decade logarithmic), lower and upper level thresholds and sample speeds. -
Page 25: Trigger
NOTICE Parameter names cannot be changed after an acquisition. Parameter names Check if the parameter names are according to your optical setup. Typical names are shown in the example above. The parameter names are displayed on the histogram axis together with a parameter label. To change parameter names, click into the fields and enter the names by using the keyboard. -
Page 26: Gain
Example: Assume vertebrate leukocytes in full blood are to be analysed. Triggering on a scatter parameter would be difficult due to the high number of erythrocytes in the same sample. Staining with a DNA dye (e.g. PI) and triggering on the DNA parameter will discriminate the leukocytes containing a nucleus from erythrocytes without a nucleus. -
Page 27: Sample Speed
The gain value of a parameter can be selected by clicking into the corresponding gain value field in the instrument settings box, please see chapter 9 Instrument Settings, Figure 7. By using the Right (Left) buttons gain values are increased (decreased). -
Page 28: Figure 9: Sample Transporting System Of The Cyflow™ Space
Typical Speed Values a) High accuracy measuremens (e.g. DNA with < 1.5% CV): Speed = 0.5 µl/s. b) Fast measurements and absolute counting: Speed = 4.0 - 10.0 µl/s Figure 9: Sample Transporting System of the CyFlow™ Space. ™ CyFlow Space | Operating Manual | March 2021... -
Page 29: Threshold: Lower Level (L-L)
9.1.4 Threshold: Lower Level (L-L) Adjustment of Lower Level L-L Setting a proper lower level (L-L) avoids the acquisition of small and unwanted background or "noise" signals below a threshold. The L-L value of a parameter can be increased (decreased) by clicking into the corresponding L-L value field in the instrument settings box and using the Right (Left) buttons, please see chapter 9 Instrument Settings, Figure 7. -
Page 30: Disposal
10.1 Disposal of device Electrical and electronic equipment must be disposed of separately from normal waste. Please contact your local Sysmex representative, since this device must be specially treated before disposal. 10.2 Disposal of components Generally, all components and fluids must be disposed of in accordance with your local regulations and acceptable laboratory procedures. - Page 31 1. Unscrew the Sheath Fluid bottle. 2. Secure Cap and Tubing on a clean surface in a stable manner. 3. Dispose of Sheath Fluid according to your local legislation and its associated labelling. 4. Screw down the Cap by turning the Bottle. Do not twist the Tubing and the Sensor cable.
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Page 32: Appendix
11 Appendix 11.1 Installation and uninstallation Installation, connection, disconnection and uninstallation may only be performed by authorised service personnel and in conformity with the applicable national rules and regulations. 11.2 Installation Requirements WARNING Overweight When moving the device (packed or unpacked) or removing it from its packaging, a person is exposed to a high risk of damaging his or her body due to the heavy weight of the device. -
Page 33: Figure 11: Instrument Setup And Periphery (Cyflow™ Space Shown From The Rear)
Instrument Setup – Overview Keyboard Keyboard Screen Screen Mouse Mouse CCD Camera CCD Camera Printer Printer Interface Interface Monitor Monitor Computer (rearside) Computer (rearside) Printer Printer ™ CyFlow CyFlow CMOS (rearside) (rearside) Computer Computer Waste Waste Sheath Sheath Waste Waste Sheath Sheath Figure 11:... -
Page 34: Instrument Setup - Step By Step
11.3 Instrument Setup – Step by Step NOTICE Before first operation remove protection material from inside the system! 1. Setting up the Computer Connect printer, screen, keyboard and mouse to the computer with the appropriate connection cables. Connect computer, screen and printer to AC power line. Figure 12: Computer connection on the CyFlow™... -
Page 35: Figure 13: Power Connection And Light Source Switches On The Left Side Of The Cyflow™ Space
Space. 3. Connecting the CyFlow™ Space to a Computer ® The CyFlow™ Space runs with a standard Windows PC equipped with a special Sysmex ® Partec interface. The integrated FloMax software controls all functions of the CyFlow™ Space, e.g. instrument settings, sample supply, sheath pressure and sample flow rate. -
Page 36: 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
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. Connect the waste fluid container to the waste outlet at the right side of the CyFlow™ Space. -
Page 37: The Flow Cuvette
Laser light exposure causes damage to skin and eyes. Avoid direct exposure. Do not open the device without authorization by Sysmex Partec. NOTICE Air bubbles usually enter the flow cuvette through the sheath fluid inlet. Bubbles might stick inside the conical section above the sample nozzle and do not enter the flow channel of the quartz cuvette segment. - Page 38 Never open the instrument to clean the flow cuvette mechanically e.g. by wires or tools for cleaning or declogging the flow cuvette. Please contact your local Sysmex representative. Removing Air Bubbles from Flow Cuvette If you notice insufficient peak resolution during sample run remove your sample and activate CLEAN several times.
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Page 39: Maintenance And Service
Put a sample tube half-ways filled with distilled water at the sample port. Clean waste and sheath reservoir, wipe top dry. 11.5.2 Service All service is to be made by authorised service personnel. Please contact your local Sysmex representative. 11.6... -
Page 40: Laser Safety
Laser light exposure causes damage to skin and eyes. Avoid direct exposure. Do not open the device without authorization by Sysmex Partec. Attention Laser radiation Class IIIb, if cover is removed and shutter is opened. Avoid eye exposure to direct laser beam. -
Page 41: Technical Data
12 Technical data NOTICE Due to fast technological improvements, specifications herein are subject to change. For details, please inquire information from your supplier. 12.1 System Size Flow cytometer: 560 mm x 650 mm x 300 mm (W x D x H) Weight approx. -
Page 42: Optics
Data Resolution 65,536 channels (16 bit) Service Please contact your local Sysmex representative 12.2 Optics Light Sources: Laser / Output Red Diode Laser: 25 mW or 40 mW at 638 nm/640 nm Laser specifications may be subject to change. -
Page 43: Fluidics
Optical Coupling Standard objective mount with high numerical aperture Excitation Optics Elliptical 15 µm x 60 µm at 488 nm NOTICE Information on the optical configuration of a specific instrument is displayed in the optical layout scheme of an instrument which can be found in the individual instruments documentation. -
Page 44: Software
Computer Desktop: Standard Desktop computer with industry standard processor, RAM, Hard Disk Keyboard and mouse Microsoft® Windows Microsoft® Office 12.5 Software ® ® 32 bit Microsoft® Windows FloMax FloMax 2.1x software routine research applications All parameters stored in FCS 2.0 or FCS 3.0 standard list-mode format Time parameter for kinetic studies Real-time acquisition... - Page 45 Protocols Individual instrument setups and acquisition displays can be saved in setup-files. The plot display, axis labels, statistics and cell concentrations are saved in documents together with the acquisition data. Report Generation Report module for Microsoft Word/Excel, ® directly called by the FloMax software: Single- or Multi-Page Report templates include institute logo(s), addresses,...
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