Re: ALP/TRAP protocols


Dr. Evans,
I believe that the enzyme methods for TRAP and AlkP will be adversely
affected by fixation and decalcification but there are IHC methods which
might work for these sections.
Patsy Ruegg

Matthew Evans wrote:

> 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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