Have a grad student who does this more than frequently, now doing it
with confocal.  Frozen sections, cut at 5 um, cold acetone fixation
for 10 min, refrig temps, air dry 30 min before fixation, fix then
air dry for another 30 min.  Staining is done (you will need to do
a dilution panel), using Dulbeccos PBS, pH 7.4 with 0.2% goat serum,
block is 5 - 10% goat serum 30 min,  staining with antibody is 30 min,
Secondary is goat antiRat F(Ab')2 biotin, 1:250 from Biosource (Jackson
has excellent secondary also), Strepavidin-HRP 1:500 from Biosource 20 min,
then chromogen of choice, we like Vector AEC or Pierce enhanced DAB.
Go easy on hematoxylin counterstain.  If you can add Tween 20 to buffer,
0.05% to 0.1% and we do both peroxidase block (Dako) and avidin biotin block)


Good luck
Gayle Callis