[Histonet] cryosectioning, bubbles between slide and section

[Christina Kreutzer]

Hello members,
I would need some help regarding cryosectioning of spinal cord. I am
currently establishing cryosectioning on the cryostat (Leica CM1950) for
this tissue and am having some problems. I am cutting 4% PFA fixed and
succrose infiltrated spinal cords, embedded in Tissue Tek at 10µm and a
temperature of approximately -16°C . I get more or less smoth slices but
once I let them adhere to the slide -directly from the knife, after
straightening it carefully with a brush - I get massive air bubbles between
the slide and the slice.

I have experience cutting on cryostats and with different tissues and never
have had this problem before.

I tried to change the temperature of the chamber and/or chunk and tried to
warm the slide before adhering the slice and I tried to cool the slide. But
it didn't help. Do you think changing from 30% succrose to a mixture of
Tissue Tek/Succrose or even Tissue Tek/PBS would help?
Does anybody have an advice?

Regards
Christina