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By the way, having read that pesky CCC article I really
wonder what data people can produce. I am amazed to see that
the bandwith between 564 and 606nm is stated to be
equivalent to a 570nm long pass in terms of what is seen of
the PE spectrum. To mention the effect of the clever angular
arangement of the dichroics in the Facsscan to change their
filter characteristics migth be too much to grasp then
anyhow. I also wonder if anyone read the article apart from
(or even not) the reviewers that it has passed without any
more comments. I haven't even heard anything with respect to
the calibration procedures.
By the way, I am also still looking for the missing 135mW of
my aircooled laser in the Elite. Anybody got them?
Gerhard.Nebe-von-caron@unilever.com
______________________________ Reply Separator _________________________________
Subject: RE: PMN Markers
Author: Kbahjat@nwu.edu at INTERNET
Date: 01/11/96 23:43
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
CD14 binding).
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...
kb
Keith Bahjat
Kbahjat@nwu.edu
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