purity with 2 colors, method for your review

From: Maciej Simm (simmmmer@yahoo.com)
Date: Thu Jan 25 2001 - 20:32:29 EST


I have a lot of fcs files from old experiments where FL3 CD45 was not
used for purity. These files include erros in lysis, nucleated RBC's,
thalassemiacs, thrombocytopeniacs and neonates (all notorious for low
purity with 2 color FACS).

I created a template which does not need the CD45 information and
extracts phenotypes for CD's 3,4,8,16+56 and 19 directly from the
fluorescence of those markers.

In my tests the results are very close to the normal analysies. But
because they're based on assumptions, there is room for error. I
created these for use in a lab that does not have 3 color capacity or
is doing retrospect analysis like me.

The assumptions are:
NK+B+T=100% lymphocytes
and
CD4+CD8= 100% CD3

anyway, I posted the template along with a bit of an overview on my
site. play around with it if it's useful and send me feedback.

I begun to post the template and description on 1/25/01, should be
ready by the next day.

http://www.cd4cd8.com/method/

maciej

PS this is work in progress :) email me if you have suggestions.



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