RE: RBC lysis problem in lymphocyte studies

From: Tom Mc Closkey (thomasm@nshs.edu)
Date: Wed Jun 23 1999 - 12:09:48 EST


--- On Fri, 18 Jun 1999 14:29:26 -0700 (PDT)  Maciej Simm 
<simmmmer@yahoo.com> wrote:

>We're experiencing consistnat difficulties w/lysing of RBC in 
>lymphocyte immunoaphenotyping. Our diff. is in the "noise" around
>the lymphocyte population, so that we can't do significant numerical
>analysis. The problem is usually conicident with young patients who
>are receiving a lot blood transfusions (such as THAL patients). We
>think that their RBC's are too strong for our lysis protocol. But at
>the same time we need to preserve the lymphocyte cell membranes b/c
>we're staining for a variety of CD markers. If anyone has any insight
>in this are please don't hesitate to share ;)

	You can add CD45 labeling to your panel and use this as a marker to 
identify the lymphs from the rbcs.  Gate on CD45+ then look at your other 
markers.

Tom


--------------------------------------------------------
Thomas W. Mc Closkey, Ph. D.
Director, Flow Cytometry
North Shore University Hospital
Biomedical Research Center
350 Community Drive
Manhasset, Long Island, New York 11030
ph: 516-562-4844 [office]; 516-562-1135/4641 [lab]
fax:  516-562-2866
6/23/99   10:12:10 AM
E-mail: thomasm@nshs.edu 
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