I forgot to soak my rat brain samples in 30% sucrose solution following perfusion and embedding in OCT compound. I am trying to section the brain, however it is quite difficult to get a good section that isn't either cracking or falling apart when I put it on the slide. Does anyone know if there is anything I can do to help with this?
Max Myers
Processing the tissue via sucrose is a dehydration step, and I think this is one of the crucial step. The tissue need to dehydrated to get the section with smooth uniform texture. Generally post mounting we expect this is irreversible step, still...Though i have not tried but i think if you should try to dehydrate the present tissue back into the sucrose, and mount it back. That may help you to find better section.
Best Wishes
Freezing the tissue without first cryoprotecting the tissue (e.g. using 30% sucrose) will severely compromise the integrity of the cells and tissue. I wouldn't count on these tissues to give useful sections.
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