DAB question

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From:Judy Trogadis <judy@playfair.utoronto.ca>
To:HistoNet@Pathology.swmed.edu (histonet)
Date:Wed, 21 Jul 1999 10:07:19 -0400 (EDT)

Dear Histonetters,

We are staining cryosections of mouse embryos which have been frozen,
sectioned and then fixed in paraformaldehyde. Immunostaining was carried 
out with the Vector Elite ABC system and the chromogen is DAB. After
the DAB reaction is terminated with water, we don't dehydrate because
we are also looking at retina which is easily damaged by alcohol, 
Therefore,we just use an aqueous mounting medium, either Ferrant's medium,  
or Aquamount. 

After coverslipping, the retinas look great. The mouse embryo DAB staining,  
however, quickly fades until it is barely visible, in a couple of hours. 
Any theories of how this could be happening?

I should add that the retinas are fixed after removal from the animal, prior
to freezing, but how could that make a difference to disolving the DAB

As usual with this group, I will be chacking for replies every few minutes.

Thank you

Judy Trogadis
Vision Science Research Program
Toronto Western Hospital Research Inst.
399 Bathurst  St.
Toronto, Canada M5T 2S8

phone: 416-603-5088
Fax:   416-603-5126
email: judy@playfair.utoronto.ca

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