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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