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" 
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=2C 
> 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=2E
> 
> 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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