April_G._Durett/MDACC%MDACC at notes.mdacc.tmc.edu
Fri Nov 1 16:32:07 EST 1996
I agree!!! We use this combination to perform a five-part diff when needed
Reference : Becton Dickinson Monograph Establishing Optimal Lymphocyte Gates
for Immunophenotyping using LeucoGATE, Michael Loken, Jeanne Brosnan, Kenneth
Ault, and Bruce Bach. The precursors will stain CD45dim CD14neg and fall in
the low SSC within the lymphocyte region and between the lymphs and mono's.
Technology Transfer-Flow Cytometry
MD Anderson Cancer Center
From: Kbahjat @ nwu.edu (Keith Bahjat) @ INTERNET
Date: 10/31/96 08:53:40 AM
Subject: RE: PMN Markers
Forgive me if I am oversimplifying, and please correct me if I'm wrong, but:
I have always had the impression that when staining a fresh peripheral
blood with CD45 FITC and CD14 PE, that a five part differential could be
performed from this 2 color plot?
With FITC (CD45) on the X axis and PE (CD14) on the Y, the Lymphocytes,of
course, are the bright CD45+, CD14 negative population. Monocytes are the
CD45 bright, CD14 bright population, with your PMNs being CD45 moderate,
CD14+(somewhat dim). The eosinophils seperate to the right of the PMNs, as
their azurphillic granules autofluoresce enough that they are brighter in
the FITC channel than the neutrophils. Basophils do not bind CD14 at all
and will sit directly below the PMNs (same CD45 intensity as PMNs, with no
As far as precursor cells (blasts, metamyelocytes, myelocytes...), I'm not
sure of their location in 45-14 plots, but have used CD45 vs SSC according
to articles published by Dr. Greg Stelzer to identify these populations. I
of course confirmed these gates by comparing with sorted cell morphology.
>From all the replies/discussion, I'm thinking I've oversimplified things?
Please correct or confirm these hypotheses...
Kbahjat at nwu.edu
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