Re: Question about biomaterial immunohistochemistry

From:Gayle Callis

If a d profile tungsten carbide knife can cut a 5 um or approx thickness,
you can remove the PMMA from the section and do immunostaining.  Contact
Neil Hand, he is an expert at using PMMA embedded tissues and
immunostaining, with many publications on technics.

If you cannot remove the plastic (very hydrophobic) immunostaining does not
work, particularly on ground sections if that is what you commonly do. 


At 11:13 AM 1/23/02 -0500, you wrote:
>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!
>Chat with friends online, try MSN Messenger:
Gayle Callis
Histopathology Supervisor
Veterinary Molecular Biology - Marsh Lab
Montana State University - Bozeman
19th and Lincoln St
Bozeman MT 59717-3610

406 994-6367
406 994-4303 (FAX)

<< Previous Message | Next Message >>