We have some sections of spinal cord from some non-human primates that my lab is trying to section. For our brain sections, we use a microtome and frozen tissue and cut 40um thick sections, not a cryostat and we are looking to do the same for the spinal cord but our first attempt was not successful. Has anyone done this successfully? We would like to avoid using a cryostat if need be as our lab prefers to have free-floating sections of tissue to work with.
Thanks!
Hi, you say "first attempt was not successful" - what went wrong? Are you cutting coronal or sagittal? Any particular area of the spinal cord, or the whole thing?
We have sectioned NHP spinal cord on a frozen stage sliding microtome. Happy to help if you are still having trouble.
Hi, you must embedded spinal cord to OCT solution then adjust temp to approximate -10 C then after sectioning place section in cryoprotectant solution (glycerol + ethylene glycol) for free-floating and OCT removement.
We find that embedding in 30% sucrose works better than OCT. This requires that you let the tissue sit in 30% sucrose until it sinks.
We have recently used the human spinal cord for immnofluorescence.
Sections were frozen and cut using a cryocut. (see attachment, Lukas et al. 2022)
A detailed protocol how to cut frozen tissue is provided in
Blumer etl al. (2019) see attachment.
We used the same protocol, for the human spinal cord
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