RE: DAB question

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From:Cynthia Favara <cfavara@niaid.nih.gov>
To:HistoNet@Pathology.swmed.edu, "'Judy Trogadis'" <judy@playfair.utoronto.ca>
Reply-To:
Date:Wed, 21 Jul 1999 12:25:25 -0400
Content-Type:text/plain

Judy,
	A few weeks ago someone had the same problem - can't remember any
solutions. what you might try is allowing the slide to air dry and then
coverslip using permount. I have used that technique when stains have faded
and it has worked well for me. Good Luck!

Cynthia Favara
Rocky Mountain Laboratories
903 S 4th Street
Hamilton, MT 59840
ph: 406-363-9317
FAX: 406-363-9286
e-mail: cfavara@nih.gov


> ----------
> From: 	Judy Trogadis[SMTP:judy@playfair.utoronto.ca]
> Sent: 	Wednesday, July 21, 1999 8:07 AM
> To: 	HistoNet@Pathology.swmed.edu
> 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: judy@playfair.utoronto.ca
> 
> 
> 



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