Re: fixated on fixation
Joseph Webster (J.Webster@centenary.usyd.edu.AU)
Mon, 27 May 1996 08:33:25 +1100
At 05:32 PM 23/05/96 -0400, Calman Prussin wrote:
>
>A question for the group:
>Has anyone had experience with losing signal after fixing previously
>stained cells? If so, with what fluorophore and what fixative? I have
>been fixing cells with 2% methanol free paraformaldehyde and seem to be
>losing my PE/Cy5 staining!
I remember that certain antigens (and certain antibodies) are killed by
paraformaldehyde fixing; I have not heard of flourochromes being affected
in the same way, but check it out anyway.
Sorry I can't find a list of affected stains, I'm sure those who work a
lot with fixed cells can provide some specific info.
The best test is simply to stain some cells, read them fresh, fix, then
repeat the reading.
Joseph.
--
Joseph Webster (O.I.C. Flow Cytometry)
Centenary Institute of cancer Medicine & Cell Biology
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