[Histonet] Help with frozen spleen and liver tissue!

[Allyse Mazzarelli]

Hi all,

Can someone please provide me with more details regarding cryosectioning of
mouse spleen and liver tissue?

Currently, I've been fixing my samples in 4% PFA (they are well fixed - I
know that will be the first question asked!), and then cryoprotect the
tissue in a series of graded sucrose solutions (15% to 30%) until they
sink.

However, when I go to place the sections on the slide from the cryostat,
they look great initially under the microscope, but once they dry they have
poor morphology, especially seen in the liver.

I've never run into this issue before, with either brain or spinal cord
which are significantly more delicate.

I've tried cutting the tissue free-floating as well to see if it was how I
was placing the sections on the slide, but nothing seems to work.

The morphology in the liver is so terribly compromised that I cannot
visualize the sinusoids properly. It is baffling that once they go onto the
slide they look okay, but 5 minutes later the tissue appears to "separate"
from itself.

Does anyone work specifically in frozen tissue sections, liver and spleen
in particular? If so, would you be able to help me figure out the best way
to generate quality specimens?

Thank you!

Regards,
Allyse Mazzarelli