MLR's with Flow

Nunez, Rafael rafaelnr at uic.edu
Tue Nov 22 10:33:41 EST 2005


WE published on Current protocols in cytometry, three protocols, basic
protocol 3, 4 and 5 that you can utilize for assessing MLR, and CTL
function. The protocols are on Chapter 9, Studies of cell function, unit
17, pages 6-11.
I can send you a copy if you do not have the CPCy book. Unfortunatelly we
dont have a PDF of the chapter, I hope it will help, REgards Rafael
On Mon, November 21, 2005 3:00 am, Guy Hermans said:
> 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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Rafael Nunez MD.
Assistant Professor of Medicine and Pathology.
University of Illinois at Chicago.
Director Cancer Center Flow Cytometry Core Facility
Room 3211 MBRB, M/C 837
900 S. Ashland ave, Chicago, Il 60607
Phone 3123550123





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