I am trying to cut 20 um cross sections of murine lymph nodes and small intestines, however, the tissue is fatty and not freezing thoroughly. When I cut the OCT, the cut will have a hole in the center where the tissue should be. Is there a better approach to fatty tissues?
I am taking the OCT directly from the -80C and the cryomicrotome is set to -20C.
Ergun Ucmakli
You can solve to use the nitrogen tank. Place the material in the holder and immerse it in a nitrogen tank with an apparatus for 3 seconds and pull the tissue at an incredibly fast speed and immediately cut the cryostat. We were able to obtain 2 micron thickness sections.
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