cryosectioning large bone samples

From:John Baker <>

Does anyone have any tips that might help me cryosection large bone 
samples?  The tissue is human navicular and cuneiform bones.  They have 
been trimmed to about 1cm thick and are about 2.5cm square. I fixed them in 
4% Paraformaldehyde for three days, placed in 10% PVA for 2 days, and snap 
frozen in isopentane in a dryice/alcohol slurry.  I have a Hacker OTF 
cryostat and fixed the tissue so I did not have contamination problems. I 
am taking 7 micron sections using the Instrumedics tape window system. But 
the tissue is not sticking to the slide but coming off with the window.  Is 
there a problem that affect the sections. Right now it just crunches into 
the bone.  Using the same spot on the knife I tested a block of trabecular 
bone and it cut like butter.  Any suggestions from those of you who cut 
bone.  Never having cut human tissues before in my cryostat what 
precautions and cleanup methods might you suggest too if I go with the 
Thank you for your time,  John

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