Re: [Histonet] c-fos staining

From:Philip Oshel



Jaina,

Most likely it's the Permount (maybe the xylenes). Use an aqueous 
mount like glycerol, methyl salicylate*, Aquamount, Fluormount, etc. 
There are others that I don't recall off hand.
After mounting, keep the slides flat. Seal with VALAP.
*Methyl salicylate is an excellent clearing medium and mountant, go 
into it using your xylenes schedule. We have specimens that have been 
in Me-sal for 14+ years and are still nice and bright. 
Rhodamine-Phalloidin stain, not slide mounted but kept in e.g. 
microfuge tubes. These aren't brain sections, but it should work for 
them, too.

Phil

>I am carrying out c-fos staining on brain tissue, and have run into a bit
>of a problem. The staining seems to work fine up untill I mount the sample
>with a xylene based mountant (permount). The stains are there after alcohol
>washing, but once I mount it, the stains seem to disappear, or become very
>very faint.
>
>Any ideas as to why this is happening, and what I can do about it.
>
>Thanks,
>
>Jaina Negandhi
>Research Project Coordinator
>Neuroscience and Mental Health
>Hospital for Sick Children
>Mc Master Building, RM 3006
>Tel: 416-813-6551
>Fax: 416-813-8724
-- 
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859

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