RE: DAB question

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From:"Nocito, Joseph" <>
To:"'Judy Trogadis'" <>,
Date:Wed, 21 Jul 1999 10:00:39 -0500

when we have to use AEC, we use Crystalmount from Fisher.  We use the
Crystalmount as a media and then place a coverslip over the top.  I have
looked back after a couple of years and the reaction was still there.  The
catalog number is BM-M02 and is about $30 for 30 mLs.  Hope this helps

Joe Nocito, B.S., HT(ASCP)QIHC
Histology Supervisor
Christus Santa Rosa Hospitals
San Antonio, Texas 

> -----Original Message-----
> From:	Judy Trogadis []
> Sent:	Wednesday, July 21, 1999 9:07 AM
> To:
> Subject:	DAB question
> 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
> precipitate?
> As usual with this group, I will be chacking for replies every few
> minutes.
> Thank you
> judy
> 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:

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