Re: IHC to see degenerative axon fibers

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From:Maria Mejia <>,

Thank you so very much for the reply. I went through my books and
finally found the protocol for this silver -Nauta-Gygax. My good-
ness the fixating time 1 week or longer (Nauta and Gygax re-
commend 1-3 months)!!! I know it says this in the protocol, but
in your experience what would be a good fixating time?? We were
hoping to perfuse w/4% PFA/0.1M phosphate buffer, then remove
the brain and fix for an additional week??? I haven't done a 
silver stain in over ten years. I want to section the tissue on
a sliding microtome (freeze the tissue after cryoprotectant) @
30um to get nice free-floating sections. The protocol says I
need to place the sections in formalin (I can use 4% PFA) - store
the sections in formalin. Can you recommend any other tips to 
this stain - if it turns out that I have to do it?? I appreciate
your time and experience - thank you.
Maria Mejia
Smith-Kettlewell Eye Res. Inst.

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