[Histonet] Re: C-fos

[Davis, Gareth]

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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