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Indeed ethidium monoazide can be used to identify "dead" cells after
fixation. Cells can be treated with a solution of EMA (concentration in
the range 1-10 micrograms/ml), the dye should enter cells in which the
membrane is damaged. It can then be photochemically stuck to the DNA by
visible light. A good reference is the paper by Riedy et al in Cytometry
(1991),12, 133-139.
EMA is available from Molecular Probes (or via Cambridge Bioscience in
the UK).
Having said all that, in our hands the technique wasnt too successful,
post-fixation the EMA positive cells lost a lot of their brightness.
Maybe others have had more luck.
Good luck anyway!
Derek
Derek Davies
Imperial Cancer Research Fund,
London, UK
http://www.icnet.uk/axp/facs/davies/index.html
On Mon, 29 Jan 1996 monard@adarc.nyu.edu wrote:
> Hi
>
> I wish to be able to identify dead cells in samples of fixed cells. The
> cells are HIV infected so they have to be fixed before running on the
> machine. I seem to remember ethidium azide can do this, has any one any
> experience of this dye? Who sells it? How do you use it?
>
> Simon Monard
> Aaron Diamond
> New York
>
>
>
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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
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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