Re: [Histonet] bone tissue falling off after AR step

From:Gayle Callis


We drain the slide, then dry bone slides flat at 37 to 40C overnight and 
longer if possible. One bonehead expert years ago cracked the bone section 
drying whip and would not dry flat any less than two weeks but that is a 
timeline you may not like.  You can then try going to a hotter oven for a 
short time, but we get more lift off of sections when the slides are dried 
in a hot oven.

Be sure the bone sections are very flat after sectioning, and that these 
little tibias are very well infiltrated with paraffin.   Good contact with 
the slide surface to begin with helps.

  At 07:56 AM 12/19/2006, you wrote:
>Hey boneheads,
>A colleague is having problems with cortical bone detaching from the
>slide during or after the antigen retrieval step. He is working with
>mouse tibia.
>He uses Superfrost+ slides. The AR is with citrate buffer (pH6),
>performed in a waterbath for 10 minutes. He bakes the slides for 15 min
>at 56C prior to deparaffinizing.
>What do you suggest?
>Thanks, Brett
>Brett M. Connolly, Ph.D.
>Research Fellow
>MRL, Imaging Research
>Merck & Co., Inc.
>PO Box 4
>West Point, PA 19486
>PH 215-652-2501
>fax. 215-993-6803
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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