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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet