For paraffin wax sections requiring Heat-induced antigen 
retrieval/unmasking, it is not often that one reads such pragmatically 
accurate, apposite and accessible statements/advice such as 
Brian's............
My only problem with it is.......
I wish I could have stated it so well!
The Researchers who carry out Immuno on "briefly" Formalin-fixed 
cryo/vibratome tissues/sections and do not need any Ag retrieval/unmasking 
to get good results should also gain knowledge from Brian's words of wisdom.
For them, I would add that when you get a poor result in such tissue prep. 
sytems, try pwax!
    An example of many such situations: In my Research Centre,  a PhD 
student  has struggled to get good results  for astrocyte subset 
localisation, using "PFA"-fixed, perfused cryo/vibratome sections with anti 
NG2. This went on for a year and then they came to me, for advice.
Once we fixed her specimens in good ol' "10% Formalin" for a week, 
pwax-processed them and carried out Immuno-DAB/Fluorochrome on the AR 
sections..beautiful results!
A year wasted!!
"Sure, "horses for courses"..
Great respect, Brian.
If I may, I will add the guts of your post to my SOPs.
Best wishes
Carl





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