Depending on the fluorphore you are using, you should mount the coverslip 
with an antifade mounting media immediately after staining is 
completed.  Do not dehydrate or clear through the same solvents as you 
would use for an H&E stain.  A good mounting media is Prolong gold antifade 
from Molecular Probes, but there are others on the market from Vector and 
others.

For a good discussion on mounting medias used for this purpose, go to IHC 
world, and into the fluorescence heading.s.

At 11:15 AM 11/4/2006, you wrote:
>I am studying tracheal vasculature by using vWF as blood vessel marker.
>   After staining i am using toluene to clear the tissue, but after using 
> that flouresence goes down..................??
>
>   Can anyone guide me for to slove this issue.
>
>   If some one have a better protocol for tracheal whole mounnt staining, 
> I would love to try for that.
>
>   Regard
>
>   Dr. Rudiger Tom
>   University of Connecticut, Storrs
>   Connecticut
>
>
>---------------------------------
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)



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