RE: Paul's Tutorial on FL1, FL2, etc.

From: Donald Mosier (dmosier@scripps.edu)
Date: Fri Dec 21 2001 - 18:09:57 EST


This is one of the few discussion groups that is worth subscribing to, and
it owes it success to the continued participation of real experts.  It is
always more interesting and illuminating when the experts don't agree.  I
thought Frank Tragonas had some interesting comments, and I eagerly await
"Zen and the Art of (Old) Cytometer Maintenance."

I am frequently called upon to review manuscripts containing flow data.  In
the past, my most frequent reasons for rejection of conclusions based on
flow data have been bad compensation, randomly set integration gates, and
no control for background fluorescence or non-specific staining.  In the
last year, I have been seeing a new problem that arises from multicolor
analysis.  Two parameter data are presented as a dot plot labeled by
antibody or reagent, but not fluorochrome (e.g., tetramer-positive cells
vs. interferon-gamma intracellular staining).  However, that data is gated
on CD3, CD8, and CD62L staining, none of which are shown.  Moreover, the
intracellular cytokine staining protocol uses monensin or some other
inhibitor of protein export which affects traffic of all surface molecules
including CD3, CD8, and CD62L.  Often no information is presented on the
fluorochromes used for these stains, but that hardly matters because the
data are not shown.

Minimal labeling of axes and display of all relevant data would make the
interpretation of flow results possible.   That now means FL1 .... FLn.

Happy Holidays.

Don Mosier
_______________________________________________________________________________
Donald E. Mosier, PhD, MD
Professor
Department of Immunology, IMM-7
The Scripps Research Institute
10550 North Torrey Pines Road
La Jolla, CA 92037, USA
858 784-9121 phone
858 784-9190 fax

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