Re: apoptosis by flow
jphillip@mednet.med.miami.edu
Thu, 04 Jan 96 09:05:26 EST
I have been reading most of the submissions on "apoptosis by flow" and
either I am missing it or no one has mentioned live gating using the
AREA AND WIDTH parameters. This is what I always do when attempting
to do DNA analysis. Putting the G0/G1 peak at a reasonable place on
the scale on an AREA histogram, and gating out doublets and very small
fragments on the area vs. width graph. If someone feels a need to use
a FSC/SSC gate you can do it on the analysis. I hope that this helps.
Jim Phillips
University of Miami (Suntan U.)
______________________________ Reply Separator _________________________________
Subject: apoptosis by flow
Author: hwortis_sup@opal.tufts.edu at smtpmed
Date: 12/27/95 7:51 PM
Is this a problem that other people have solved?
We have been using flow to analyze apoptosis in lymphocytes. We
have been using P.I. staining and using either ethanol or hypotonic lysis
to obtain nuclei and stained nuclear fragments. The problem is whether
to use FSC/SSC gates. If the answer is yes, which particles do you omit
from the gate?
It seems that there is a real problem of analysis. Each
apoptotic nucleus may form several fragments. Therefore there
is no defined relationship between the number of fragments and the number of
apoptotic cells. Do other flowers recognize this as a problem? If so
how do you solve it?
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