Optimizing PMT Gains on Digital Cytometers

Stingley, Robin L StingleyRobinL at uams.edu
Mon Jan 23 09:56:37 EST 2006


I've tried it two more times, and got spikes at different voltages each
time.  The third time, I got two spikes in the same run.  I'm trying to
use them all to get an average for the correct PMT settings.  I don't
really understand what's happening.  

 

Robin

 

Robin Stingley, Ph.D.

Flow Cytometry Core Facility

Department of Microbiology and Immunology

University of Arkansas for Medical Sciences

Slot 511, Rm B504

4301 West Markham Street

Little Rock, Arkansas 72205

Phone:	501-686-6927

E-mail:  StingleyRobinL at uams.edu

http://www.uams.edu/flowcytometry/ <http://www.uams.edu/flowcytometry/> 

 

________________________________

From: Carol Oxford [mailto:cloxford at ucdavis.edu] 
Sent: Thursday, January 19, 2006 6:55 PM
To: cyto-inbox
Subject: Re: Optimizing PMT Gains on Digital Cytometers

 

That's really interesting- I've done it twice now, and both times my
curves look very similar to yours, although several of the parameters
that started out with lower CV's didn't show the spike.   I was about to
send the data to BD for some advice- I'd be really interested to see
what data other labs are getting.

 

Carol

 

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Carol Oxford					       

Manager, UC Davis Optical Biology Lab	    

Tupper Hall, rm 3425				 

University of California			  

Davis, California  95616		        

(530) 752-7205					    

(530) 752-4548 fax				   

cloxford at ucdavis.edu  

http://ccresources.ucdmc.ucdavis.edu	        

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 On Jan 17, 2006, at 4:09 PM, Stingley, Robin L wrote:





Dear Flow Experts,

 

 

 

Thanks so much for all the help on my questions about using auto comp

and the other features of FACSDiva.  Many of you mentioned that I should

establish the optimum baseline PMT gains using the BD application note

for digital cytometers to help with the rare and dim populations of

cells that we are trying to analyze.  I've just tried the protocol, and

my performance curves look weird.  I've attached a PowerPoint slide of

them.  There's a spike in the %CV of each fluorescence parameter at PMT

voltage 600.  

 

 

 

Should I continue on, or is there some kind of problem either with the

way I set this up or with the instrument?   

 

 

 

Thanks,

 

Robin

 

 

 

Robin Stingley, Ph.D.

 

Flow Cytometry Core Facility

 

Department of Microbiology and Immunology

 

University of Arkansas for Medical Sciences

 

Slot 511, Rm B504

 

4301 West Markham Street

 

Little Rock, Arkansas 72205

 

Phone:	501-686-6927

 

E-mail:  StingleyRobinL at uams.edu

 

http://www.uams.edu/flowcytometry/ <http://www.uams.edu/flowcytometry/> 

 

 

 

 

 

 

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This attachment - 'Baseline settings 011706.ppt' -  62.46 KBytes - can
be viewed at

http://www.cyto.purdue.edu/MD-parts/d6f3c3be0d8e0560a89c59259895ecf0070e
9e40.ppt 

 

 

 

------------------------------------------------------------------------
---

Carol Oxford					       

Manager, UC Davis Optical Biology Lab	    

Tupper Hall, rm 3425				 

University of California			  

Davis, California  95616		        

(530) 752-7205					    

(530) 752-4548 fax				   

cloxford at ucdavis.edu  

http://ccresources.ucdmc.ucdavis.edu	        

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---

 

 


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