RE: bone IHC

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Date:Thu, 03 Jun 1999 09:56:35 -0600
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Not much published specifically for bone, that I am aware of, we are
pioneers in this area.  We have been doing some ISH on frozen bone sections
using the Instrumedics Tape Transfer method, just about impossible without
it in my experience.  Formic acid decalcified paraffin embedded zinc formal
fixed is how I do IHC on bone samples, and if any pretreatments are required
I use enzyme digestions rather than HIER.  If you would like to join the NSH
IHC Resource Group send me a fax number and I will fax you an application.
The group has a general IHC reference list available to our members.
Patsy Ruegg 

		-----Original Message-----
		From: []
		Sent:	Wednesday, June 02, 1999 12:25 PM
		Subject:	Re: bone IHC


		I am just beginning to set up our lab for IHC and in situ on
bone - frozen and
		paraffin.  Do you have any recommendations for reading
materials and any helpful
		hints?  I'm sure there are many but a few to get me started
would be helpful.  I
		have never fixed, processed or cut bone frozen or in
paraffin - only when
		embedded in MMA.  This is quite a stressful thing to start
when I don't have any
		experience.  Thanks for any suggestions you may have.

		Deb Ebert
		Procter & Gamble Pharmaceuticals
		Mason, OH

		From: on 06/02/99 12:53 PM

		cc:    (bcc: Deb Ebert-DC/PGI)
		Subject:  bone IHC

		I missed the original thread on Bone IHC, so now I would
like to add my two
		cents worth.

		I too have gone to fixing with Zinc formal which requires
much less antigen
		retrieval, but when I do require pretreatment to access the
antigen site, I
		use enzyme digestions rather than hier.  We have found that
we can control
		the process much better with enzyme digestion and it causes
less damage and
		dislodging of the sections.  I also use silane coated slides
(plus slides).
		There are certain components in bony tissues that require
enzyme digestion
		anyway and you must completely decalcify or the hydroxy
appetite will block
		access to the antigen site, or take on everything because it
is so porous
		and absorbent.  Before IHC I always treat the deparaffinzed
slides with 0.4
		M EDTA to assure complete decalcification.  There are so
many idiocyncrities
		about bone IHC, it is a whole different science from soft
tissue IHC.
		Patsy Ruegg


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