[Histonet] Trouble with AcP staining MMA sections

[eplurbus <@t> u.washington.edu]

Hello,

I am having trouble staining some old (7 years) undecalcified, ethanol fixed, methylmethacrylate embedded bone for the presence of acid phosphatase. The blocks that I am trying my protocol on were successfully sectioned and stained by a former technician about four years ago but I haven't been able to duplicate her results. I am using a paraffin embedded cat tooth as control and the osteoclasts stain red every time, but I get no response from the MMA embedded sections. I've tried it after deplasticizing, not deplasticizing (trouble keeping the sections on the slides), re-hydration, without re-hydration....

I feel that my protocol is good and should allow for the detection of AcP but perhaps somewhere along the line I am not handling the MMA embedded tissue appropriately.

Is there anyone out there with experience on staining methylmethacrylate embedded tissue for the presence of acid phosphatase that may have a suggestion? I would be happy to post a detailed account of my procedure for someone with experience in these matters.