Dear colleagues,
I would like to cryosect aortic plus perivascular adipose tissue from obese mice. Unfortunately, at -23°C, with a new knife, embedded in Tissue Tek and very careful handling, the tissue is too unstable and it tears when cutting.
Thicker cuts have shown no improvement.
Do you have any tips on how to cut the tissue better?
Also maybe someone who has only cut fat tissue?
I am very glad for help:)
Hi Jasper,
Do you cryoprotect your samples prior to embedding in OCT with sucrose gradient? We routinely fix our mouse aortic samples in 4% PFA O/N, 15-30% sucrose gradient, followed by embedding in OCT compound in cryomold, and freezing with liquid nitrogen-cooled isopentane.
Also, you can refer to the following thread on fat tissue embedding/sectioning:
https://www.researchgate.net/post/How-to-get-cryosection-from-oily-organs-like-adipose-tissue
Hope that helps!
Thanks for the tip. Until now, I have not fixed the tissue in succrose before embedding.
I will try that and also cut in colder temperatures:)
Kind regards
Jasper
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