zinc fixation/EDTA

From:Joey Shepherd

Afternoon all!

I'm going to try zinc fixation (fixer from Pharmingen) on some mouse 
limbs for 24-48hr, then decalcify in 20%EDTA. I usually decalc. after 
formalin fixation but the antigens I'm after (CD4, CD19..) aren't 
liking that, hence the zinc. However, I have a question about this 
before I go ahead- will the EDTA somehow strip the zinc as well as 
the calcium from the sections, or will it be stable after the 
fixation step? My boss has raised concerns about this. He also seems 
to think that "EGTA" is an excellent calcium chelator with a lower 
avidity for clacium - does anyone use this instead of EDTA?



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