RE: HIER on bone tissue

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Efrain and interested others:

We do various post-processing techniques on bony tissues. We Fix and
dehydrate the tissues then clear with Methyl salicylate. This seems to
significantly decrease the brittleness and facilitates cutting. We cut a 5
microns and place the sections on BD Gold Seal Ultrastick or Polysciences
SectionLock slides. These slides work best for our special staining.
Finally, I heat the retrieval buffer in the microwave to the desired
temperature THEN place the slides in the hot liquid for the required amount
of time. We let them set on the bench-top at room temperature for fifteen
minutes. No problems with the bone falling off. Bob Skinner is giving his
bone processing workshop at NSH this fall and I'll be giving a follow-up
workshop on special techniques in bone.If you can't make the workshops we
can probably send you the handouts after the meeting.

Good luck, Donna Montague

-----Original Message-----
From: Efrain Pacheco []
Sent: Thursday, August 03, 2000 10:14 AM
To: Histonet
Subject: HIER on bone tissue

Does anybody has any procedure to do HIER (heat
induced epitope retrival)on bone tissue without
loosing the cortical bone? I have two antibodies that
appear to work well after microwave HIER except for
the fact that the hard bone comes off the slide,
apparently because of the heat. By the way, we are
working with rat femurs. I will appreciate any

Efrain Pacheco, HT(ASCP)
Histology Associate
Amgen Inc. 

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