Re: PMT Cascade Experiment

Hello Ck,

hope this is what you ask about:
over past two ISAC meetings Jo Trotter of BD gave presentation about 
setting your PMT above your resolution threshold.
See their publication Maecker, H. T., and Trotter, J. (2006). Flow 
cytometry controls, instrument setup, and the determination of 
positivity. Cytometry A.

My simplified version of the same test is:
Use Rainbow 8 peak beads (we use them for daily performance control), 
start at PMT voltage at 300 and increase by 50 in all detectors, record 
data with each PMT value.
Then you can simply go through the data and see the threshold PMT where 
you start to see the most peaks (usually 8 peaks for FITC, PE detector, 
6-7 peaks for further red, 7-8 peaks for violet laser excited detectors).
That PMT value is your resolution threshold. You should not use PMT 
lower then that or you loose some resolution (in your experiments 
resolution of low intensities might be hampered by autofluorescence, but 
it is a separate issue)

Hope this helps Tomas

Ck Poon wrote:
> Hello All,
>
> We are trying to test the sensitivity of some PMTs on our LSRIIs and 
> ARIA. If I remember correctly, Stephen Perfetto mentioned about a 
> cascade experiment for doing so at ISAC2006. I was wondering if any of 
> you know how this experiment/test is performed. Any guidance will help.
>
> Many thanks,
> Ck
> ++++++++++++++++++++++++++++++++++++++++
> Ck Poon
> Department of Experimental Medicine
> University of California, San Francisco
> UCSF Box 1353
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>
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-- 
MUDr. Tomáš Kalina, Ph.D.
Laboratoř průtokové cytometrie - CLIP
Klinika dětské hematologie a onkologie
Universita Karlova - 2. lékařská fakulta

Skype: tomaskalina
fax (+420) 22443 6521 (6417)
http://www.lf2.cuni.cz/clip/
tel: (+420) 22443 6477
adresa: 	V Úvalu 84
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***********************************************

Tomas Kalina, M.D., Ph.D.
Flow Cytometry Lab - CLIP
Department of Paediatric Hematology/Oncology
2nd Medical School  
Charles University Prague

Skype: tomaskalina
fax (+420) 22443 6521 (6417)
http://www.lf2.cuni.cz/clip/
phone (+420) 22443 6477
postal address:       V Uvalu 84
		      150 06 Praha 5
		      Czech Republic