decalcification and cryostats

From:jerry hu <>

Hello Everyone,

I have been through the histonet archives and found
many recipies for bone decalcification, and have
looked at some books on this topic.  However, I have
also noticed that bone is generally processed through
paraffin, though what I have avaliable is a cryostat -
are there major differences that I should be aware of?

What I plan to do is fix my specimen (3 mm thick) for
24 hours in formalin, then decalcify in buffered
formic acid (pH ~2), and do endpoint determination
with ammonium oxalate.  Then I would like to
freeze-embed the decalcified specimen in freeze
embedding compound and section on a cryotome.  Does
the freeze embedding medium need to infiltrate the
specimen (as is the case with paraffin processing) to
cut well?  

What I would like to do is to find the thickness of
the cartilage above the bone with H/E.  Any comments
or experience on this will be most appreciated.  I
would be *really happy* if there is a way to get
decalcifying the bone.  Thanks!

Jerry Hu
Rice University
Houston, Texas, USA

PS sorry about the generic yahoo address - my school
e-mail system does not allow me to post to Histonet -
kinda like the problem John Kiernan is facing.

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