Reporting results of 2-colour stains

From: Paula Lavery (plaver@po-box.mcgill.ca)
Date: Mon Dec 06 1999 - 13:03:48 EST


Hello Everyone out in Flow-Land

Back to Flow Cytometry 101...

I've been having a debate with our grad student on the appropriate way of
reporting results of 2-colour staining. He is staining activated T cells
with a PE-conjugated activation marker and another FITC-conjugated
monoclonal. He wants to make a statement on Activated vs. Non-activated
cells. He prints up contour graphs with both colours and calls the UR
quadrant his "FITC+ on activated cells" and the LR quadrant his "FITC+ on
non-activated cells". I think he should be gating on activated cells (with
FL2 histogram) and then produce an FL1 histogram to get the "FITC+ on
activated cells". (Similar with non-activated cells, of course.) The numbers
are substantially different depending on the method you use. Help please!

Paula Lavery
Transplant Immunology Lab.
Royal Victoria Hospital
McGill University
Montreal (QC) Canada
plaver@po-box.mcgill.ca

"If we knew what we were doing it wouldn't be called research, would it?"
-- Albert Einstein



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