From: /G=Gerhard/S=Nebe-von-Caron/OU=1890CHPE/O=TMGB.URC/@LANGATE.gb.sprint.com
Date: Tue Aug 29 1995 - 04:04:00 EST
Don't remember who asked, but problems of aggregation
between Leukocytes are nicely visible when looking at linear
side scatter versus log fluorescence. CD3 positive
granulocytes can be immediately spotted (Detection of
Leukocyte cell-aggregation and its effect on light scatter
differentiation of lymphocytes, Cytometry 1994, Suppl.7,
page 64).
Looking at aggregates in the lymphocyte scatter cluster only
remains problematic because of the variation, blast cells
and the scatter position for different cell types in that
cluster as pointed out by Leon WMM Terstappen in his Thesis.
You also have to consider the problem of coincidence down to
flow (event) rate and garbage that is gated out when on its
own, but still can come through the gate together with a
cell.
Good luck
Gerhard Nebe-v.Caron
Unilever Research, Colworth Laboratory Sharnbrook,
Bedfordshire
GB - MK44 1LQ
Tel: +44(0)1234-222066
FAX: +44(0)1234-222344
E.mail: gerhard.nebe-von-caron@urcgb.sprint.com
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Subject: Lymphocyte Clusters
Author: Dennis_Young@cis.ucsd.edu at INTERNET
Date: 26/08/95 19:37
Using the DDM (Doublet Discrimination Module) can help, but
cells are not perfect little billard balls, so it won't
*EVER* be 100% accurate. The other problem is that conjugates
are rare, even when you try to make them!
The simplest way is to look at scatter signals. Cell clusters
will have higher Forward and Side scatter signal intensity.
See the recent postings on doublet disc in DNA analysis.
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* Dennis J. Young Voice : (619) 822-0407 *
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* University of California, San Diego USA e-mail: djyoung@ucsd.edu *
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Our lab uses a BD FACScan and performs three-color analysis on
human periphal blood lymphocytes. Does anyone know of the best
way to identify lymphocyte conjugates (clusters of two or more cells)?
Tim Patton
University of Pittsburgh
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