For thawed cells, PI and Fsc should be very reliable for excluding dead and dying cells. If PI can get in, so can your antibodies, leading to artefacts. ======================================================= Rachel M. Gerstein, Ph.D. Department of Molecular Genetics and Microbiology Graduate Program in Immunology/Virology University of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655-0002 (508) 856-1044 (508) 856-5920 (FAX) > ---------- > From: Bergh, Ole > Sent: Tuesday, February 5, 2002 3:09 PM > To: Cytometry Mailing List > Subject: RE: staining for viability > > 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: Cytometry Mailing List > 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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