Hi everyone,
I have some questions about performing an immunohistochemical stain on 
PMMA-embedded rabbit skull bone containing a hydroxyapatite and/or composite 
biomaterial.  One of my concerns is whether the blade that we have to cut 
samples will cut through the biomaterial.  Has anyone ever had problems 
using a Reichert-Jung Polycut E microtome to cut PMMA?  Also, I would like 
to stain for either alkaline or acid phosphatase.  The embedding process was 
done at room temperature until the end when it was polymerized at 37 degrees 
C in a vacuum oven.  My practice samples are all embedded in PMMA with 2.5% 
benzoyl peroxide.  Will the enzyme activity have been destroyed?  I am also 
concerned if the tissue within the biomaterial will stain, if the 
biomaterial is dense like hydroxyapatite.  I would like to not have to 
decalcify the sample since the biomaterial remaining in the sample is pretty 
important to me.  If ANYONE has any ideas at all, please let me know.  I am 
working blind here, I have done lots of reading but I wanted to know if 
anyone else had any ideas before I started.  Thanks!

Tithi


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