I think that the vertebral column might damage the blade or making it completely impossible. Has anyone ever tried to do this?
Hi Elena,
Yes i have quite good experience in sectioning spinal cord after perfusion. I was just wondering if would have been possible to do the same but with the columns still in place. HAve u ever tried?
Thanks very much.
Matteo, yes it is possible, you have to decalcify the bones. There are good reagents for that used in many anatomo-patology labs. We did it a couple of times and it gives decent results even if they are never optimal.
Hi Matteo, without decalcification you can't cut the vertebrae in a cryostat. Quick decalcification protocols normally use acids which is not very good to the spinal cord, longer decalcification protocols with e.g. EDTA might work reasonably ok, but in my opinion the sectioning and the staining will never be as good as if you would process the spinal cord alone. My question is why would you need to do that?
Agree, the decalcification protocols may lead to reduced spinal cord histologic quality. If the issue is to register the spinal cord and vertebral segments, then MRI imaging of the vertebrae/cord first will greatly help interpret the spinal cord slides.
Hi Elena,
I am now trying to cut a mouse spinal cord at the cryostat. I have been using the vibratome, so far but I am not satisfied with the quality of the sections. What is your protocol to cryopreserve the sample/
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