Re: [Histonet] C-fos stain

From:

Geoff McAuliffe

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

Davis, Gareth wrote:

I plan to do a immunohistochemical stain on mouse brain tissue with c-fos. Some protocols I've read state that you should osmicate the sections, after mounting, to enhance the stain. I've never stained with c-fos, but I have been told it is very dark on it's own.=C2� Does anyone know if it will be necessary to osmicate. Of course, I guess I could just try and see, but any information on the staining process itself would be helpful.

Thanks,

Ms. Gareth Davis

Vanderbilt University Medical Center

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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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 
mcauliff@umdnj.edu
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