RE: DAB question
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From: | "Nocito, Joseph" <joseph_nocito@srhc.iwhs.org> |
To: | "'Judy Trogadis'" <judy@playfair.utoronto.ca>, HistoNet@Pathology.swmed.edu |
Reply-To: | |
Date: | Wed, 21 Jul 1999 10:00:39 -0500 |
Content-Type: | text/plain |
Judy,
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 [SMTP:judy@playfair.utoronto.ca]
> Sent: Wednesday, July 21, 1999 9:07 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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