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CyFlow
space
Instrument Operating Manual

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Summary of Contents for Partec CyFlow space

  • Page 1 ® CyFlow space Instrument Operating Manual...
  • Page 2: Table Of Contents

    Laser Safety ..............................23 ® CyFlow space - Technical Specifications ....................24 For in vitro diagnostic use with Partec recommended IVD reagents. ® The Partec CyFlow space flow cytometer complies with the European IVD Directive 98/79/EC and is therefore marked with the CE sign.
  • Page 3: Introduction To Cyflow Space

    Introduction to CyFlow ® Introduction to CyFlow space What... ® … is the Partec CyFlow ® space? The Partec CyFlow space is a fully equipped desktop Flow Cytometer (FCM). It features a modular optical concept which allows using up to three different laser light sources and between one and nine optical channels (parameters).
  • Page 4: Typical Steps Of Particle Analysis

    1. Preparation and Staining Typical Steps Particle For flow cytometric analysis, the cells (or other Analysis particles) must be separated from each other in aqueous suspension. Typically, generating sufficient optical signals themselves, the cells are then labelled by staining with more fluorescent dyes.
  • Page 5: Operating Basics

    The display shows empty histograms. 6. Cleaning Use speed 4. Run 1.6 ml Partec Cleaning Solution until sample tube is empty. Activate CLEAN. Run a second tube with Partec Sheath Fluid for 2 minutes. ®...
  • Page 6: Multi Laser Measurements

    The waste may contain biohazardeous and cancerogenic material from the samples (infectious cells, dyes). To minimize biohazards, fill about 25 ml of Partec Hypochlorite Solution 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 throughoutly with soap and disinfectant.
  • Page 7: Starting A Measurement

    830 µl for true volumetric absolute settings. counting and not more than 2.8 ml. For smaller amounts of sample use Partec “Small Volume Sample Tubes” (Product no. 04-2010, however these tubes are not compatible with the True Volumetric Absolute Counting option, page 10).
  • Page 8: Switching Off The Cyflow Space

    15 minutes. Re-start the system by pressing START and let it run to the end. Run the system with 1.6 ml Partec Cleaning Solution until the sample tube is empty. Run 1.6 ml Partec Sheath Fluid for two minutes. Press STOP. 2. Protect the sample port Leave the final sample tube connected to the sample port –...
  • Page 9: True Volumetric Absolute Counting - Overview

    All Partec instruments volume (Please contact Partec or your local are specifically designed to minimize counting distributor for details). losses by providing direct connection between 9/26 ®...
  • Page 10: Performing True Volumetric Absolute Counting

    Partec CountCheck beads (Product no. 05- 4010). iii) Less Preparation or Setup Time The sample concentration is directly analysed by...
  • Page 11: Optical Standard Setup - The Parameters

    Operating Basics Optical Standard Setup – The Parameters ® The CyFlow space flow cytometer can be Note equipped with various laser light sources (1, 2 or 3 If required by a specific application, the optical lasers), different laser power intensities and up to standard setup can be optimized by exchanging nine optical parameters.
  • Page 12: Optical Flow Geometry

    Optical Flow Geometry waste side scatter fluorescence forward scatter laser beam (hydrodynamic focusing) sheath fluid sample Fig. 3: Optical geometry at the flow cuvette (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 13: The Parameter Setup Dialog Box: Pulse Height, Area And Width

    Operating Basics The Parameter Setup Dialog Box: Pulse Height, Area and Width Before starting acquisition, assure parameters of interest are selected and set up according to your analysis. Opening the Click "Parameters..." button Parameter Setup Instrument Settings box. The Parameter Setup Dialog Box appears. Fig.
  • Page 14: Trigger

    Leading Trigger Trigger In order to discriminate particles of interest e.g. cells from other particles e.g. cell fragments or nutrition particles in cell culture, a proper trigger ® has to be chosen. In the CyFlow space, any parameter can be used as leading trigger. Frequently, the forward scatter parameter (FSC) or, preferently for smaller particles, the side scatter parameter (SSC) is used to trigger on all...
  • Page 15: Pmt High Voltage, Gain, And Log Amplification

    Operating Basics PMT High Voltage, Gain, and Log Amplification Controls Typical effect Adjustment change Gain By means of the gain, the signal amplification can be individually adjusted in a wide range for each parameter. For the used photomultiplier tubes (PMTs) by changing the gain value, the PMT's high voltage is adjusted.
  • Page 16: Sample Speed

    Operating Basics Sample Speed Speed Typical effect of Adjustment change By means of the speed value, the sample speed (in terms of µl/s) is set. The speed value can be increased (decreased) by clicking into the corresponding field in the instrument Note settings box and using the Right (Left) buttons (see EAT cells, Speed...
  • Page 17: Threshold: Lower Level (L-L)

    Operating Basics Threshold: Lower Level (L-L) Lower Level (L-L) Typical effect of Adjustment change 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 L-L = 10 (decreased) by clicking into the corresponding L-L value field in the instrument settings box and using...
  • Page 18: Appendix

    Appendix Installation Requirements Line Power ® CyFlow space Basic Unit AC 100-240 V 50-60 Hz, 250 VA max Instrument Size ® CyFlow space Basic Unit 560 mm x 650 mm x 300 mm (W x D x H) Operating Environment ®...
  • Page 19: Instrument Setup - Overview

    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 (rearside) (rearside) Computer Computer Waste Waste Sheath Sheath Waste Waste Sheath Sheath ®...
  • Page 20: Instrument Setup - Step By Step

    Note: Before first operation remove protection Instrument Setup – Step by Step 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.
  • Page 21: The Flow Cuvette

    The laminar flow of sheath fluid and 3. Press START and wait till automatic stop sample flow as well as hydrodynamic focusing are 4. Place a tube with 1.5 ml Partec Cleaning disturbed and broad peaks appear in the Solution on the sample port and repeat histograms.
  • Page 22: Maintenance And Service

    Wash the flow cuvette by contact your local distributor or Partec. attaching a tube with 1.6 ml of Partec Sheath Fluid and run it for at least 2 minutes. Maintenance and Service ®...
  • Page 23: Laser Safety

    Appendix Laser Safety ® The CyFlow space is a class I laser product according to the EN 60825-1 : 2001. Warning: Laser light can be emitted if the protection cover for the laser beam is removed and the beam shutter is opened.
  • Page 24: Cyflow ® Space - Technical Specifications

    Appendix ® CyFlow space - Technical Specifications Note Due to fast technological improvements, specifications herein are subject to change. For details, please inquire information from your supplier. ® 1. CyFlow space System Size Flow cytometer: 560 mm x 650 mm x 300 mm (W x D x H) Weight ca.
  • Page 25 Appendix Warranty 12 months on all parts except filters, mirrors, other quartz or glass parts, disposables and cuvettes ® 2. CyFlow space Optics Laser / Output Red Diode Laser: 25 mW or 40 mW at 638 nm Green solid-state Laser: 30 mW to 100 mW at 532nm Blue solid-state Laser: 20 mW or 200 mW at 488 nm...
  • Page 26 Appendix ® 5. FloMax Software ® 32 bit Windows™ FloMax Software software for routine and research applications All parameters stored in FCS 2.0 or FCS 3.0 standard listmode format Time parameter for kinetic studies Realtime acquisition Multiparameter N-colour compensation (online and offline) Multiparameter gating, MultiColor gating Peak and cluster analysis and statistics DNA-cell cycle analysis...

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