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Introduction to CyFlow

Rev. 007 Date 2010-01-13

CyFlow

space Instrument Operating Manual

2/26

Table of

contents

Introduction to CyFlow

space ....................................................................................................................... 3

Typical Steps of Particle Analysis .................................................................................................................. 4

Operating Basics .............................................................................................................................................. 5

Switching on the CyFlow

space ................................................................................................................. 5

Multi laser Measurements .............................................................................................................................. 6

Starting a Measurement ................................................................................................................................. 7

Switching off the CyFlow

space ................................................................................................................. 8

True Volumetric Absolute Counting

– Overview ............................................................................................ 9

Performing True Volumetric Absolute Counting ........................................................................................... 10

Optical Standard Setup

– The Parameters .................................................................................................. 11

Optical Flow Geometry ................................................................................................................................. 12

Instrument Settings ....................................................................................................................................... 12

The Parameter Setup Dialog Box: Pulse Height, Area and Width ............................................................... 13

Trigger .......................................................................................................................................................... 14

PMT High Voltage, Gain, and Log Amplification .......................................................................................... 15

Sample Speed .............................................................................................................................................. 16

Threshold: Lower Level (L-L) ....................................................................................................................... 17

Appendix ......................................................................................................................................................... 18

Installation Requirements ............................................................................................................................. 18

Instrument Setup

– Overview ....................................................................................................................... 19

Instrument Setup

– Step by Step ................................................................................................................. 20

The Flow Cuvette ......................................................................................................................................... 21

Maintenance and Service ............................................................................................................................. 22

Transport and Storage.................................................................................................................................. 22

Disposal ........................................................................................................................................................ 22

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.

IVD

Содержание CyFlow space

Страница 1: ...CyFlow space Instrument Operating Manual...

Страница 2: ...ent Settings 12 The Parameter Setup Dialog Box Pulse Height Area and Width 13 Trigger 14 PMT High Voltage Gain and Log Amplification 15 Sample Speed 16 Threshold Lower Level L L 17 Appendix 18 Install...

Страница 3: ...lly any flow cytometric application The applications cover e g Routine and research Immunophenotyping Blood Cell Analysis HIV monitoring Leukocyte Counting Rare Event Analysis Microorganism Analysis F...

Страница 4: ...haracteristic 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 cel...

Страница 5: ...ometer is operated with 100 240 V AC Switch on main power at the left side of the instrument The 488nm laser has its own power button closeby please also switch on the 488 nm laser All other lasers ar...

Страница 6: ...s of the flow cuvette The CyFlow space Flow Cytometer supports either 2 laser spots when employing FloMax version 2 6 or 2 7 or 3 laser spots when employing FloMax version 3 0 The blue 488 nm laser wh...

Страница 7: ...nsert sample tube onto the sample port until you recognize a click The sample should be fully mounted within a second Now the measurement acquisition starts automatically the operating software indica...

Страница 8: ...ath fluid tube for 5 seconds Stop the instrument while keeping the tube pinched 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 Parte...

Страница 9: ...ential This requires fast recognition and analysis of the events by electronics and computer All Partec instruments are specifically designed to minimize counting losses by providing direct connection...

Страница 10: ...No additional analysis steps e g setting gates for beads are required v Less Expenses No reference beads required Performing True Volumetric Absolute Counting Sedimentation and Count Time Cells or oth...

Страница 11: ...rward scatter SSC side scatter FL1 green fluorescence FITC FL2 orange fluorescence PE FL3 red fluorescence I PE Cy5 FL4 far red fluorescence I PE Cy7 FL5 red fluorescence II APC FL6 far red fluorescen...

Страница 12: ...ion 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 le...

Страница 13: ...meter label To change parameter names click into the fields and enter the names by using the keyboard For each optical channel you may analyse three different pulse properties 1 The pulse height 2 the...

Страница 14: ...l other particles will not be recognized by the instrument The trigger parameter can be used for an efficient exclusion of unwanted particles from the analysis Example Assume vertebrate leukocytes in...

Страница 15: ...the instrument settings box see Fig 4 By using the Right Left buttons gain values are increased decreased In case linear amplification is used peaks are expanded to the right compressed to the left w...

Страница 16: ...decreased by clicking into the corresponding field in the instrument settings box and using the Right Left buttons see Fig 4 The count rate increases with elevated speed values If the speed is increas...

Страница 17: ...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 setting...

Страница 18: ...educe smoke dust vibrations direct sunlight and direct neighbourhood of heatings as possible The installation room must be well ventilated and dry Temperature 15 30 C Humidity 20 85 relative non conde...

Страница 19: ...space shown from the rearside CyFlow rearside Screen Computer rearside Printer CCD Camera Interface Monitor Printer CCD Computer AC AC AC Air Sheath Mouse Keyboard Sheath Waste AC Waste CyFlow rearsid...

Страница 20: ...e supply sheath pressure and sample flow rate The Computer connections of the CyFlow space are located at the rear side of the instrument Connect the CyFlow space to the computer with the interface ca...

Страница 21: ...during sample run remove your sample and activate CLEAN several times In this way the flow cuvette is back flushed with sheath fluid Restart the sample and observe peak resolution again If the results...

Страница 22: ...lled water and a clean brush and flush with clean destilled water several times Remember cleanliness of sheath fluid reservoir is critical for proper operation If the CyFlow space will not be used for...

Страница 23: ...1 2001 Warning Laser light can be emitted if the protection cover for the laser beam is removed and the beam shutter is opened Therefore the system is marked with the following laser safety labels War...

Страница 24: ...entation control Particle Concentration Analysis True Volumetric Absolute Counting Particle Size and Fluorescence Distribution Analysis True Volumetric Based on precise counting and mechanical fluid v...

Страница 25: ...d precision syringe pump for contamination free sample transport Built in air pressure for sheath fluid Sheath fluid pressure is adjustable from 0 300 mbar Computer controlled Default setting 200 mBar...

Страница 26: ...d to 32 gates in free logical combinations Regions quadrants 1P ranges Gating Crosstalk On or offline gating and crosstalk compensation provide adjustments without need Compensation to rerun samples D...

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