RE: decalcification and cryostats
You could get around bone decal by using the Instrumedics tape transfer
system. It may be difficult to cut fixed samples in the cryostat, some
people do it by rinsing out the fix and decal I would imagine very well with
buffer and infiltrating the sample with sucrose. John Tarpley could help
you with this.
From: jerry hu [mailto:email@example.com]
Sent: Wednesday, March 21, 2001 4:43 PM
Subject: decalcification and cryostats
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
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!
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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