[Histonet] cryosectioning, bubbles between slide and section

[Emily Brown]

Hello,

This may be a little late, but we use a mix of half 30% sucrose and half
OCT to section.  It works really well with 14 to 25 micron sections and no
cooling or heating of the slide.  Our tissue is very small
though--embryonic spinal cord or brain from chick or mouse.  I just change
the way I pick it up to get rid of bubbles.  Have you tried rolling the
slide upwards when you pick it up (if that makes sense) as opposed to
rolling it from left to right (or the opposite)? Also as our cryostat gets
older, it tends to get more static and we cannot get rid of it.  This
causes the tissue to adhere to the slide faster, which definitely causes
bubbles.  That may the case, but I hope not!!

Emily

"By bitching and bitching and bitching, they could exhaust the drama of
their own horror stories. Grow bored. Only then could they accept a new
story for their lives. Move forward."

-Chuck Palahniuk, "Haunted"


On Fri, Apr 25, 2014 at 2:58 AM, Christina Kreutzer <
christina.kreutzer01 <@t> gmail.com> wrote:

> 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
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