[Histonet] Plastic processing for undecalcified mouse bones

[Collette, Nicole M.]

Hello, All,

I am looking into starting a plastic embedding/sectioning endeavor, and am trying to figure out my startup reagents and protocols. I am seeing a lot of protocols for the Technovit system, which uses a slide press to adhere the sections to the slides, and would prefer to avoid expensive startup equipment if possible. I found another protocol that uses silanized slides, water and heat to adhere the sections. Is this comparable to results seen with a slide press, or is there some obvious down-side? I have been mainly looking into MMA/PMMA protocols for my bone, and ideally would like to be able to use the sections for antibody stains in addition to histology stains and mineral assessment- although I will work with the limitations of the medium, I know I can’t always ask for the moon. I will be using adult mouse bones, primarily from appendicular skeleton (long bones). I am trying to start with a relatively do-it-yourself, low throughput option that minimizes startup cost for a system that I may only use short-term. Up until now I have been using either paraffin embedding (decalcified samples), or frozen Cryojane sections (unfixed, undecalcified), but there is potential for plastic to be the best option in some instances. I think I have the sectioning capabilities covered, but would be appreciative of embedding and sectioning protocols (and sage advice from the wise, experienced bone cutters out there, if I’m totally headed in the wrong direction!). A catalog# recommendation for molds/chucks/cassettes (to fit or otherwise adapt to a Leica RM2255 microtome) would be fantastic. Thanks in advance for your help and support.

Sincerely,
Nicole Collette
Lawrence Livermore National Lab
collette2 <@t> llnl.gov