Re: Freeze whole blood or buffy coats??

From: Candace Enockson (enockson@musc.edu)
Date: Tue Aug 31 1999 - 08:53:41 EST


Stan, I have a user who routinely freezes his cells prior to staining for
surface markers and cytokines. However, he does not freeze the whole blood
as you asked.  He processes the blood as he would for non frozen samples -
stimulates, lyses, fixes - then suspends them in 10% DMSO and 90% FBS and
freezes.  When he's ready to run, he thaws and stains. Works fine.

Candace Enockson
Medical University of South Carolina  

--On Fri, Aug 27, 1999 5:33 PM +0000 "Ress, S, Stan, Dr"
<sress@uctgsh1.uct.ac.za> wrote:

> 
> Hi Folks,
> 
> This question has been posted before by myself and others, but I'm 
> still in the dark, so here I go again...
> 
> Does anyone have a protocol for freezing whole blood that 
> would permit subsequent flow cytometry on the thawed sample? We have 
> a study starting but the ab have not all arrived.
> 
> Alternatively, is there any data/personal observations on comparison 
> of frozen buffy coats vs ficoll gradient PBMNC with regard to flow 
> cytometry? There must be some information on this, someone must have 
> tried it!
> 
> Any data or advice would be greatly appreciated.
> 
> Thanks,
> 
> Stan



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