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.
Patsy Ruegg

		-----Original Message-----
		From:	jerry hu []
		Sent:	Wednesday, March 21, 2001 4:43 PM
		Subject:	decalcification and cryostats

		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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