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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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