Re: PI gating on FL3

Dave Coder (dcoder@u.washington.edu)
Wed, 30 Jul 1997 16:39:37 -0700

Yes, if the PE overlap into the FL3 detector is compensated properly (you
can, of course, ignore the very bright PI into FL2). If you draw the
exclusion gate on the PE vs. PI dot plot the proper boundary may be more
clear. See the web page below for an example with 7-AAD.

http://nucleus.immunol.washington.edu/Research_facilities/Apps/7aad.html

Dave Coder
dcoder@u.washington.edu

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From: pathol-immuno biol-ther <biol-ther.pathol-immuno@psl.ap-hop-paris.fr>
To: cyto-inbox
Subject: PI gating on FL3
Date: Wednesday, July 30, 1997 3:09 AM

Dear all,
I need your comments to settle a discussion in the lab regarding PI-based
gating of viable
cells in FL3. I do membrane staining with a combination of FITC and PE
conjugated
Moabs, then I stain cels with PI, make exclusion gate on PI-positive cells
in FL3 and then
evaluate FL1/FL2 staing on PI-negative cells. Is that a correct approach
and if not what are
other ways?
Thank you,
Karolina Palucka