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Appendix 

 

Rev. 007 Date 2010-01-13

 

   

CyFlow

®

 space Instrument Operating Manual 

21/26

 

 

The Flow Cuvette  

 

 Note 

The  flow  cuvette  is  “the  heart”  of  your  flow 
cytometer.  It  is  responsible  to  guide  cells  and 
particles  through  the  center  of  the  illumination 
spots with a micrometer precision. The flow cell is 
manufactured  with highest  possible  precision  and 
should  be  treated  like  that.  Any  deformation  will 
degrade  the  flow  precision  and  make  it 
irrepairably unusable. 
 

 

 
 
 

Fig. 12: Flow cuvette with sample, sheath fluid 
and waste connections. 
 

 Note 

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.  The  laminar  flow  of  sheath  fluid  and 
sample flow as well as hydrodynamic focusing are 
disturbed  and  broad  peaks  appear  in  the 
histograms. 

 

 Note 

Do not readjust the laser or laser optics.  
 

 DANGER 

Warning: Do not look into the laser beam. Use 
eye  protection  glasses  of  filters  or  block  the 
laser beam. 
 

 

Cleaning of the flow cuvette 

 
For 

cleaning  the  flow  channel

  use  Partec 

Cleaning Solution

 (Product no. 04-4009) filled into 

a  standard  sample  tube  and  connect  to  the 
sample  port  2-3  times.  Then  perform  additional 
cleaning  cycles  with  destilled  water  filled  into  a 
fresh sample tube. 
Do not use mechanical wires or tools for cleaning 
or declogging the flow cell. 
 

Removing Air Bubbles from Flow Cuvette 

 
If  you  notice  unsufficient  peak  resolution  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  are  still  insufficient,  run  Partec  Calibration 
Beads  (e.g.  3  µm  Calibration  Beads).  Compare 
the  result  with the  exemplary  data files  located  in 
the  FloMax/QC  or  FloMax/Calibration  data 
directory  and  observe  the  flow  monitor  (CCD 
camera). A single bright spot should appear in the 
centre of the visible field. If this is not the case air 
bubbles  or dirt  inside the flow cell may cause the 
disturbance.  As  first  easy  approach  to  release 
bubbles 

during  system  run

  attach  a  tube  with 

Partec  Cleaning  Solution  and 

pinch  first  the 

sheath and then the waste tubing 

several times 

to  remove  air  bubbles  through  the  flow  channel 
into the waste. 
 
In  case  air  bubbles  are  still  persistent  please 
apply the following procedure: 
 

1.  Select the instrument settings: 

Sample prerun = 15 sec 
Stabilizing time = 5 sec 
Automatic stop = 15 sec 

2.  Place  an 

empty 

sample  tube  on  sample 

port 

3.  Press START and wait till automatic stop 
4.  Place  a  tube  with  1.5  ml  Partec  Cleaning 

Solution  on  the  sample  port  and  repeat 
step 3. 

5.  Reset  instrument  settings  to  normal 

values  

 
If analysis with Partec Calibration Beads does not 
result  in  acceptable  signal  distributions  an 
incubation 

of 

the 

flow 

cell 

with 

Partec 

Decontamination  Solution  is  required.  Run  the 
system  with  1.6  ml  Partec 

Decontamination 

Solution

.  During  system  RUN  pinch  the  sheath 

fluid tube for 5 seconds. Stop the instrument while 
keeping  the  tube  pinched  by  pressing  STOP. 

Waste outlet 

Quarz cuvette 

Sample nozzle screw 

Sample inlet 

Sheath fluid inlet 

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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