autofluorescence

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
To:histonet@pathology.swmed.edu
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One can try antifade mounting medias from Molecular Probes and often
coverslipping with pure PBS (seal the coverslip with diluted mounting
media, fingernail polish contains isopropyl alcohol that can mix with
aqueous mounting media i.e. PBS) will work.   

There are substances that autofluoresce in cells that cannot be quenched,
and after some discussion here, it was pointed out the if our fluorescent
signal was brighter than the autofluorescence, one may be able to live with
it. 

We went to a brighter fluorescent dye (brighter signal, less quenching) the
Alexa dyes from Molecular Probes, and are not bothered by the low level of
autofluorescence in the tissues.

In April and October, there was a great deal of discussion about
autofluorescence/mounting media etc.  Go back into the Histonet Archives,
you will find some excellent discussions on this problem.






Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303



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