I need a pressing question answered. I am working with someone that
insists that fixation and decalcification should be done at degrees for ISH.


We are fixing mouse tibia in 4% PFA for 48 hours at 4 degrees and decaling
in 10% EDTA for 14 days at 4 degrees both on a rocker with several changes.

I use formalin for 48 hours at RT and 10% EDTA for 14 days at RT for similar
projects and have no problems. The stuff for ISH is dry, brittle and hard to
cut. 

Is it really necessary to do fixation and decalcification in the refrig? I
am at my wits end!!

Thanks

Jennifer Harvey
University of Rochester