3 color phenotyping

Dawson, Carolyn cdd1 at CIDHIV7.EM.CDC.GOV
Thu Feb 10 09:07:00 EST 1994


I have tried to use PerCP on my FACstar Plus and have had similiar results.  
While attending the BD Symposia in San Francisco, I spoke with an engineer 
who explained that the problem with PerCP is that it photo-bleaches 
more rapidly in the stream-in-air instrument.  I was able to get a very dim 
signal compared to the FACscan.  As a result I was unable to use this 
fluorochrome.

Carolyn Dawson 
 
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Subject: 3 color phenotyping
To: cytometry at flowcyt.cyto.purdue.edu
Date: Mon, 7 Feb 1994 15:34:54 +0200 (WET)
From: Yashpal Agrawal <yagrawal at messi.uku.fi>
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Dear Dr. Langweiler,
We tried 3 color analysis on an FacStar using PerCP derivatives. No 
fluorescence was observed in the Fl3 channel for the weakly positive antigens,
e.g. CD19. We contacted the manufacturer, who suggested that we change the
standard filter set to another which was more suitable. On changing the filter
set, still no fluorescence was seen. We were then told that PerCP cannot be
detected on our FACSstar, as it was not sensitive enough, since apparently
perCp can be detected on a FACScan. I dont remember clearly the situation for
the brightly staining cells. As I remember even those antigens were only dimly
fluorescent in Fl3.

It would be interesting to know whether PerCP derivatives can be measured on 
a FACStar, our machine is from the year 1988/89.
-- 
Yash Pal Agrawal MD, PhD
Dept. of Clinical Chemistry,                     Tel:+358-71-173155
Kuopio University Hospital,                      FAX:+358-71-173200
POB 1777, 70211 Kuopio Finland                  E-mail: yagrawal at messi.uku.fi







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