|From:||Bret Morrow <firstname.lastname@example.org>|
Sorry, nothing to add on the 'flowery twats.' However, I'd like to ask
what the conventional wisdom is on using fetal tissue in
immunohistochemical (IHC) staining for tyrosine hydroxylase (TH) using a
monoclonal. I have played around with it a bit, but still have a few
1) How long do people immersion fix fetal tissue after transcardiac
perfusion with 4% paraformaldehyde? I have seen 1-3 hrs and several
weeks for IHC, but not for the TH antbody specifically.
2) I have tried egg yolk embedding then freezing microtome cutting
and am not wild about it, although the yolk does help hold the fetal
tissue together. I have read about gelatin embedding--any experience?
--any other options?
3) Finally, my usual peroxide/methanol quenching of endogenous DAB
reactivity left the tissue in a sorry state. I am currently planning on
skipping this step, but as I want to count dopamine neurons, I need to
avoid the nasty red blood cells. Any thoughts?
Bret A. Morrow, Ph.D.
Associate research scientist
Associate clinical professor
Yale University School of Medicine
New Haven, CT 06520-8066
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