MLR's with Flow

Guy Hermans Guy.Hermans at ablynx.com
Mon Nov 21 04:00:44 EST 2005


Hi Don,

Have you tried going for a CFSE-dilution type experiment? Assuming you first
purify the T-cells, say on a MACS, I'd stain them with a FACSarray
compatible CFSE analogue, mix with unlabeled APC's and try to observe
downward shifts in fluorescence intensity.

CFSE won't work on the Array, but I've got two non-overlapping spectra dyes
working already and a third one waiting to be tested. Mail me directly if
you want the gory details.

Guy

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-----Original Message-----
From: Wigington, Don [mailto:Don.Wigington at genzyme.com] 
Sent: Friday, November 18, 2005 12:12 AM
To: cyto-inbox
Subject: MLR's with Flow


Dear Flowers

Here is a pretty simple question. (But I bet not a lot of simple answers.)

Has anyone ever tried to use flow cytometry for analyzing mixed leukocyte
reactions? I am trying to study the proliferation of responder T cells rate
mixed with drug treated APC's. We can't use radioactivity here so I was
going to try a colorimetric approach (cell titer 96;Promega) but the
sensitivity is not great enough. I am a bit worried that I will not have
enough cells for flow but I do have a BD FACSArray so I can do it in 96-well
plates. Any info would be appreciated very much. 

Don

-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-
Don Wigington
Manager, Preclinical Laboratory
Genzyme Corporation
1600 Aspen Commons
Middleton WI 53562
608-662-7866  fax 608-662-7870


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