Processing Intervertebral Disks
Hello Histonetters,
I am having some problems processing en block intervertebral disk specimens
(flanked by portions of vertebrae). I am especially interested in
examining the nucleus pulposus. The samples are about 1 cm thick, fixed
for about 10 days in 10% formalin and then decalcified in 5% formic acid,
anywhere from 1 to 1.5 weeks. The processing schedule I am using is 1.5
hours in each of the dehydrating, clearing, and paraffin stations. After
coarse trimming to the area of the nucleus pulposus, I am finding that this
portion of the paraffin block is soft like is was improperly infiltrated
and/or processed. For the most part, the anulus fibrosus is intact except
at the nucleus pulposus interface.
Is this simply a matter of increasing the processing times or is there
something else I am neglecting to do? Any help would be greatly appreciated!
Gratefully yours,
Tracey
Tracey Couse
Laboratory Coordinator/Histologist
Georgia Tech/Emory Center for the
Engineering of Living Tissues
Georgia Institute of Technology
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