RE: [Histonet] Bone adhesion problems after HIER

From:"Liz Chlipala"


Its very difficult to maintain section adherence of bone with HIER
techniques.  We try to use enzyme digestion methods over HEIR when ever we
can. What antibody are you looking at?


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309

-----Original Message-----
[] On Behalf Of Leung,
Sent: Thursday, January 11, 2007 10:18 AM
Subject: [Histonet] Bone adhesion problems after HIER


I am having problems with cortical bone detaching from the slide during
the antigen retrieval step. The samples I am staining are paraffin
embedded mouse tibia.

I am currently using Superfrost+ slides. The AR is with citrate buffer
(pH6.1) from Dako performed in a waterbath for 15 minutes. The slides
are baked overnight at 59C prior to deparaffinizing. 

I have tried using a steamer and microwave, however I still ended up
with the same results. In addition Superfrost GOLD slides did not help
in keeping the bone on. I found that performing HIER at 70C for three
hrs as suggested in the archives to be ineffective in exposing my
antigen (although the bone did stay attached). 

Do you have any suggestions?


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