[Histonet] placental IHC for the novice

[Julienne Rutherford]

Hello. I'm new to histonet and am not trained in histo or IHC, but am very
interested. I'm throwing myself into the fray, and hope you can help me. In
response, which I welcome privately or in the forum, please use simple
language for I am but a simple woman with a simple dream of studying monkey
placentas.  I want to study IGF-1 localization in the marmoset monkey
placenta. I'm doing a lot of reading regarding methods, but I need help as
to what I need to do (and by extension, how to ask for the money to do it in
grant applications!). The folks in our histo lab are going to help me and
train me but I want to come to them more prepared and I have to provide the
specific materials I'll need. I have both 10% NBF-fixed tissues (most
sitting around in formalin for a very long time), and I also have frozen,
but by this I mean stuck in the regular old lab freezer to freeze
conventionally. I will begin snap-freezing placentae we collect from now on,
but what can I get out of this old stuff? Can I thaw out what I have and
snap-freeze them in OCT? Should I try to do IHC on both the formalin-fixed
AND the frozen tissue?

In order to build a budget I need to know everything I need to make these
slides. So, what kind of slides and coverslips do I need, what are all the
chemicals involved, what is a secondary antibody and why do I need it, is a
chromagen different from a stain, etc.? Also, is there something akin to and
"IHC for Dummies" resource? I've looked at ihcworld.com for protocols but
it's over my head at this point. I recognize that this is a skill perfected
over many years, but I really would like a primer on the basics so I can
have a more informed discussion about it.

If the elementary pitch of my questions is inappropriate to the forum, I
sincerely apologize. I'm just not sure where else to turn. Please respond to
my Texas e-mail if you'd prefer to school (or scold) me privately!

Thanks in advance,
Julienne Rutherford

Research Assistant (Adjunct)
Southwest Foundation for Biomedical Research
Southwest National Primate Research Center
P.O. Box 760549
San Antonio, Texas 78245-0549
phone (210) 258-9864
fax (210) 258-9883
jrutherford <@t> icarus.sfbr.org

Ph.D. candidate
Indiana University
Bloomington, Indiana
jnruther <@t> indiana.edu