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 ---------- 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
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