Fetal tissue

From:Bret Morrow <bret.morrow@yale.edu>


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?

TIA, y'all!

Bret Morrow

Bret A. Morrow, Ph.D.
Associate research scientist
Associate clinical professor
Yale University School of Medicine
New Haven, CT 06520-8066

voice 203.785.4515
fax  203.785.7670
email  bret.morrow@yale.edu

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