[Histonet] Bone Cracking During PMMA Embedding

[Rachel Paietta]

Hello,

       Our research group has been using a PMMA embedding protocol to embed
bone samples and bone/soft tissue samples, but we have noticed recently in
our BSE SEM images that we are getting an unacceptable amount of cracking.
I am looking for some advice or any suggestions to prevent this and what
other people have done if they have had a similar experience.  We believe we
are probably having some issues with the dehydration steps, but are unsure
about what to change or what may be causing the issue.  The procedure we use
is as follows:

Dehydration in Ethanol - 70% > 70% > 80% > 95% > 95% > 100%> 100%
Acetone
Monomer 1 (MMA, Benzoyl Peroxide and Dibutyl pthalate)
Monomer 2 (MMA, Benzoyl Peroxid, Dibutyl Pthalate and PMMA beads)
Monomer 3 (same as monomer 2 with more PMMA beads dissolved)

Thanks in advance for your help.  I am also posting a BSE image of one of
our samples to illustrate the cracking.  The file
is:"PMMAembedded_sheep_spine_cracking.jpg" and is taken at 30x.

Sincerely,
Rachel

-- 
Rachel Paietta

PhD Candidate, Mechanical Engineering
427 UCB, University of Colorado at Boulder
Boulder, CO 80301
paietta <@t> colorado.edu