RE: staining for viability

From: Bergh, Ole (berghoj@MSX.UPMC.EDU)
Date: Tue Feb 05 2002 - 15:09:13 EST


Fluorescein diacetate will stain live cells green, ethidium bromide will
counterstain red for dead cells. For further details:

Takasugi, M. An improved fluorochromatic cytotoxic test. Transplantation
1971 Aug 12(2):148-51

Ole J. Bergh, Supervisor
Flow Cytometry Facility
University of Pittsburgh Cancer Institute
200 Lothrop Street  W 1009 Biomedical Science Tower
Pittsburgh, PA 15213
Phone:412 624 0399  Fax: 412 624 9624
mailto:berghoj@msx.upmc.edu <mailto:berghoj@msx.upmc.edu> 
http://pci.upmc.edu/internet/flowcyto/flow.htm
<http://pci.upmc.edu/internet/flowcyto/flow.htm> 


-----Original Message-----
From: Michal Abel [mailto:abelm@necker.fr]
Sent: Friday, February 01, 2002 3:43 PM
To: cyto-inbox
Subject: staining for viability



I've got a question for the list regarding staining for viability.
I am working with freesed/defrost lymphocytes. One of the drawbacks
of this procedure is the number of death cells, which make higher the
background noise. I wonder what kind of marker should I use for
gating only the alive lymphocytes. Logically, I thought, the best
would be necrosis marker like PI. However, I am not sure if I also
need to eliminate the early apoptotic cells or not ?
I would be grateful if someone could help me resolving this problem.

Thanks,


--
Michal Abel, MD, PhD
Laboratory of Clinical Immunology
INSERM U25
Necker Hospital, 161 Rue de Sèvres. Paris
tel: 33 1 44 49 52 23
fax: 33 1 44 49 53 74



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