[Histonet] histology of rat knee joint for TRPV1, CGRP, SP

[Saniya Fayzullina]

Hello,

I am trying to work out a protocol for immunohistochemistry on the rat
knee. I have the following questions:

1. Is decalcification necessary? How long is enough in EDTA? I will
probably embed in OCT, and cut on a cryostat, so I am assuming the bones
might create a problem. 

2. Do 20um sections sound reasonable for the joint? 

3. I would prefer to stick with a simple
block->primary->fluorophore-conjugated-secondary protocol, rather than
ABC, and dealing with streptavidin etc steps. Has anyone successfully
used a TRPV1 receptor antibody on rat tissue? CGRP? SP?

4. What do you think about the formalin-picric acid perfusion method?
Does the picric acid really make a big difference? I have seen it used
at 0.2%, 2%, and 15%. Which is preferred?

Thanks!

~Saniya