Hello!
I have just encountered a new issue with some tissue flaking off of my slides and wondered if anyone else has experienced this or has any suggestions? The sections were cut at 18 microns in a -19 degree cryostat and were thaw-mounted on superfrost plus slides. When the tissue was dried, it began to flake off the slides. The brains were perfused with 0.01M PBS and 4% PFA, then post-fixed in 4% PFA for 24 hours before being allowed to sink in 30% sucrose (~3 days).
If anyone has encountered this issue or has any suggestions, please leave a comment! Thanks
hi Reneta, i dont know ur research subject, but i can recommend u a basic technic from pathology laboratory froze section routine. After taking section from tissue, u can immerse the preparate into 96% ethylene alcohole to not flake off
Dear Renata, it looks like the tissue was not well fixed and lost water after the sectioning, which caused it to flake off. In my past experience, I have fixed perfused rat brains for up to 72h in pfa 4% and used a sacarose gradient (10 - 20 - 30%) instead of a single 30% sucrose solution, which is hypertonic and may have promoted that water loss. We used the samples for immunohistochemistry and the morphology was intact. Hope it helps.
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