Author Topic: Mystery artifact in brain tissue!  (Read 744 times)

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Offline LHelou

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Mystery artifact in brain tissue!
« on: December 08, 2016, 01:45:15 PM »
Hello all,

Thanks for helping me think through a problem I'm having.  I'm cutting 50-micron frozen sections of an adult macaque (fascicularis) brain. As is our lab's protocol, the brain sat in formalin + 10% glycerin for a couple days before it was extracted from the skull.  I then extracted it and put it into formalin + 20% glycerin for 20 days (we typically do at least 14).  I embedded the brain in a mixed medium of OCT and glycerin, then froze it.

When I first began sectioning the brain at the frontal pole, the tissue simply disintegrated, or at least was badly mangled. I stopped cutting, melted down the block, and put the brain back into formalin + 20% glycerin for 5 days, hoping to cryoprotect it further.  When I refroze the brain and started cutting again, it cut better than previously, but still not good enough to continue.  There are artifacts in the tissue that resemble those caused by a nick in the blade, but (A) I'm using a newly sharpened blade, and (B) the lines are not in the same location across sections.

I don't have pieces of pia hanging off the tissue surface, I don't have debris on the blade, there are no nicks on the blade, I can't see ice crystals in the tissue, I've cryoprotected the tissue for twice as long as usual...what else should I be looking for?  Calcium deposits (this was not an old animal)? Any other things I can try to salvage this case?

Thanks in advance!

Mystery artifact in brain tissue!
« on: December 08, 2016, 01:45:15 PM »

Offline GAlison

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Re: Mystery artifact in brain tissue!
« Reply #1 on: February 08, 2017, 01:21:30 PM »
I've been advised to never unfreeze and refreeze tissue. I've had tissue disintegrate once I put it into the well so my solution for that was to directly mount the slice onto the slide. How are you fixing the tissue?

Re: Mystery artifact in brain tissue!
« Reply #1 on: February 08, 2017, 01:21:30 PM »