Greetings,

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
Q's:

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