I fix the bones in 4% PFA in PBS at 4C for 48 hours. Then I place the bones
in 0.5M EDTA (disodium salt) pH=8.0 and keep them for 10-20 days at 4C
rocking and changing the EDTA every 3 days.
Then I do 30% sucrose and OCT embedding. 20 micron is very thick try to cut
Directly conjugated secondary is better and easier if it works! If it does
not you may try the other options......
I think personally that picric acid is niot good idea to fix for IHC, unless
the antibody does not work with regular fixatives and is reported to work
with picric acid.
Memorial Sloan-Kettering Center
New York, NY 10021
Histonet mailing list