nissl stain in primate tissue-problem

Erin eme11 at psu.edu
Thu Mar 6 13:02:18 EST 2003


Hello! I was wondering if anybody could help me to figure out what is
going wrong in my staining. We are processing primate tissue that has
been injected with HRP. It is fixed with 4% paraformaldehyde (made
fresh). One series of the tissue is to be stained with cresyl violet,
but during the stain the tissue becomes damaged/dry and curls up off
the slide on the edges and around and injection/lesioned sites. (The
series is not reacted for HRP, just mounted and air-dried overnight).
This same tissue stains fine with neutral red. And the cresyl violet
protocol works for tissue that is fixed with 10% formaldehyde (no
HRP). The CV protocol I use does include 5 mins in choloform, is that
too harsh for this tissue? This is the CV protocol we use: 100% EtOH,
chloroform (5 mins), 100% EtOH, 95% EtOH, 70%EtOH, dH2O, CV, dH2O, 70%
EtOH, 95% EtOH, 95%+acetic acid, 95% EtOH, 100% EtOH, 100% EtOH,
xylene (all of the above are just for a few dips except for the CV
stain and the differentiation fluid). It is a protocol I was given -
is there a better protocol for CV staining? Any insight into my
problem would be greatly appreciated! Thanks!



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