PI/DNA plus PE and FITC staining
Dr. Howard Petrie (h-petrie@ski.mskcc.org)
Wed, 21 Aug 1996 19:21:38 -0400
Dear Flow Users: I am trying to develop a working method for two-color
surface staining plus DNA content analysis on a FACScan. In my hands, none
of the fixation/permeabilization methods that maintain surface
immunofluorescence generate adequate 7-AAD histograms (high CVs). I have
been concentrating on trying FITC/PE plus PI (ca. 0.5-1 microgram/ml). I
can compensate PE/PI at this level, and CVs are adequate, although
certainly not as good as PI at higher concentrations. The problem I appear
to be having is that (apparent) PE fluorescence disappears when PI is
added. I'm sure it's some compensation dilemma, but I can't seem to make
headway with this problem. Does anyone use such a method, and if so, can
you suggest a compensation protocol? I will repost a summary in the event
of multiple responses. Thanks for your help,
Howard Petrie, Ph.D.
Assistant Member, Immunology Program
Memorial Sloan-Kettering Cancer Center
Box 341, 1275 York Avenue
New York NY 10021
phone (212)639-2149
fax (212)794-4019
e-mail h-petrie@ski.mskcc.org
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