ALP/TRAP protocols

From:Matthew Evans

Hi all
Two questions, bit long-winded, so apologies.

I'm looking at samples of cranial bone around an implant made of
polycaprolactone polymer and am currently approaching it in two ways.
1) 4% PFA fixation, formic acid decalcification, cryosections.
2) Ethanol fixation, embedding in Technovit 7100 Glycol methacrylate
resin (Methylmethacrylate resins melt the implant).
-
Does anyone have a protocol for detection of alkaline phosphatase and
tartrate-resistant acid phosphatase that could be applied to either of
these two section types ??.
I believe the decalcification and formalin fixation may make detection
in the cryo-sections difficult.
-
Is it likely that we'll be able to cut thicker sections of the Tecnovit
7100 samples on a diamond saw for SEM purposes (2mm thick) ??.
Or is this resin just too soft to support bone plus implants.

Thanks for any help in advance

Matt

Dr Matthew Evans
Research Associate
School of Biomedical Sciences
University of Nottingham
E-Floor Medical School
Queens Medical Centre
Nottingham
UK
NG7 2UH





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