[Histonet] C-fos stain

[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.  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
>  
>  


-- 
--
**********************************************
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 <@t> umdnj.edu
**********************************************


-------------- next part --------------
An HTML attachment was scrubbed...
URL: http://lists.utsouthwestern.edu/pipermail/histonet/attachments/20030909/c4d5438e/attachment.htm