Frozen sectioning via microtome, sections 40 um thick.
I am wondering if it looks like the sections are thinner in those ventral, whiter spots (perhaps from the tissue warming too much in that spot?)--or, perhaps, a dehydration artifact, if perhaps these sections were sticking out a bit from the cryo solution in the tube or during washes. These two sections are 10 sections (400 microns) apart from each other. Or a freezing artifact -- but other sections look fine.
I notice the very edges of sections similarly turn opaque white sometimes as they dry. Maybe these points are slightly lifted off the slide?
Thank you for your help!
Thank you for sharing this Dr. Hepker. I presume it may be a focal area of incomplete dehydration prior to clearing and mounting. Try using 100% methanol or add acetone as a secondary dehydrating agent prior to clearing with xylene or benzene. Please update me if this resolves the problem.
Tnx for sharing. same problem occurred for me and this problem is about section lift off on edges and I found when section placed on gelatinated lam some times OCT solution thawed under section and disrupt section adherence to gelatin so after dehydration this part of section not adhered to lam. for solve this problem I remove OCT solution from section and make sure nothing placed between lam and section.
Thank you so much, it could be some debris and air bubbles, as well as an adherent of gelatine particles
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