[Histonet] Re: C-fos
Luz Cardenas
lcardenas <@t> utmem.edu
Wed Sep 10 15:22:24 CDT 2003
Hi Geoff,
I am also trying C-fos in brain tissue, free floating and I would like to try nickel-cobalt recipe is you don't mind, can you send it to me also?.
Thanks!
Luz Cárdenas, MD
Post-doctoral Research Trainee
Department of Anatomy and Neurobiology
University of Tennessee Health Science Center
855 Monroe Avenue, Room 309/302
Memphis, TN 38163
Lab: (901) 448-6002 / 7318
Fax: (901) 448-7193 (Dept)
----- Original Message -----
From: "Davis, Gareth" <gareth.davis <@t> Vanderbilt.Edu>
Date: Wednesday, September 10, 2003 3:04 pm
Subject: [Histonet] Re: C-fos
> Geoff,
> Thanks for the information on osmium. I have used the stuff for
> Scanning EM and know how awful it is to use, so would love to
> avoid it. I was going to try staining free floating sections with
> c-fos, but have decided to mount the sections onto glass slides
> first. We have done nickel enhancement when staining cell
> cultures, and decided it didn't make much of a difference. But,
> maybe your nickel+cobalt recipe would be helpful for our sections.
>
> Thanks,
> Gareth
>
>
>
>
> Geoff replied to C-fos and osmication question:
> Hi Gareth:
>
> I have not seen an answer to your question on Histonet so
> ......... Osmium (1% aqueous or less) will enhance the DAB
> reaction product. If your (brain) sections have not been
> defatted/delipidized osmium will also stain the lipids (myelin) in
> the brain. This may actually reduce the contrast of the DAB-osmium
> reaction product depending on where it is. Osmium will also make
> the sections less flexible so they should be mounted on slides
> before exposure. Osmium is expensive and must be used under a hood
> ("fume cupboard") as its vapors will fix your corneal and nasal
> epithelium. It is also requires some care in its disposal. There
> are several less dangerous/expensive ways to enhance the DAB
> reaction product, including nickel or nickel+cobalt ions in the
> DAB-peroxide incubation medium is popular. While nckel and cobalt
> are certainly not benign they don't have the fume problems that
> osmium does and they are less expensive. I can send you a
> nickel+cobalt recipe if you like. In my experience DAB should be
> 'enhanced' with something if you want the intensity of the
> reaction product to be permant. Also, use DPX as a mounting media.
>
> Geoff
>
>
> Ms. Gareth B. Davis
> Research Assistant II
> Neuro-magnetics Division of
> the Department of Neurology
> Vanderbilt University Medical Center
> 615-936-3318
>
>
>
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