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Operating Basics 

Rev. 007 Date 2010-01-13

 

   

CyFlow

®

 space Instrument Operating Manual 

7/26

 

 
 

Starting a Measurement 

 

 

 

 Notes 

 
See  FloMax

®

  -  Acqui-

sition  and  Instrument 
control  manual  on  how 
to  change  instrument 
settings. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
When  removing  a  tube 
from  the  sample  port, 
the BioSafety system is 
activated 

to 

prevent 

dropping 

and 

contamination 

by 

hazardeous 

material. 

Simultaneously, 

cleaning 

cycle 

is 

performed 

to 

avoid 

cross-contamination 
between 

individual 

samples. 
 
 

 
 
 
 
 

 

 
 
 
 
 

 

 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 
 
 
 
 
 

 

or

  

 

 
 

 

 

 

or

  

 

 

1.

  Make  sure  your  flow  cytometer  is  ready  for 

analysis  and  the  operating  software  is  prepared  for 
the measurement. 
 

2.  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  Partec  “Small  Volume 
Sample  Tubes”  (Product  no.  04-2010,  however 
these  tubes  are  not  compatible  with  the  True 
Volumetric Absolute Counting option, page 10). 
 

3.  Insert  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  indicates the 

Prerun

Stabilize

Run

 

and 

Count

 

status. 

Instrument status is also highlighted by LEDs on the 
front side of the CyFlow

®

 space. 

 

 In the 

Prerun

 phase, cells are quickly transported 

to the position of analysis, the flow cuvette. 
 

  During  the 

Stabilize

  phase,  the  system  waits  a 

given time for the flow to reach slow speed required 
for analysis. 
 

 In the 

Run

 phase, cells are analysed and classified 

into histograms on the display in realtime. 
 

  In the 

Count

  phase, cells  are  counted  for  a  given 

volume.  Do  not  change  instrument  settings  in  the 
count phase. 
 
After  the  count  phase,  the  acquisition  finishes 
automatically.  To  finish  the  acquisition  before  this 
manually, click the end icon or end button, or simply 
remove the sample tube from the sample port. 
 

 The BioSafety cycle is started. 
 Status changes to 

Clean

 and then 

Ready

 
To  restart  an  acquisition  without  removing  the 
sample tube, click onto start. 
 

4. To save results, click on the disk icon. 

 
 

 
 

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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