Live/dead cell discrimination
ZBIGNIEW DARZYNKIEWICZ (darzynk@nymc.edu)
Sat, 16 Nov 1996 15:18:19 -0500
Live vs. dead cells can be discriminated, after fixation, based on
the differences in DNA sensitivity to denaturation. Both, apoptotic
and necrotic cells have DNA highly sensitive to denturation, which
can be probed treating cells with 0.1 M HCl and staining with
acridine orange (6 - 8 ug/ml) at pH 2.6. The method is described e.g.
in our chapter entitled "Measurements of Cell Death by Flow
Cytometry" in "Techniques in Apoptosis", a User's Guide series. T.G.
Cotter and S.J. Martin, Eds. Portland Press, London, 1996, pp 71-106.
The problem with this approach, however, is that DNA in mitotic cells
also is unstable and stains similar as in dead cells. One has to
correct, therefore, for the possible presence of mitotic cells in the
sample.
Zbigniew Darzynkiewicz
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
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PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
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