[Histonet] Frozen sectioning human spleen

[James S.]

I've been trying to get some frozen sections of human spleen. Usually all the work I do is with mouse tissue and I take the tissues myself and immediately freeze in OCT in isopentane/dry ice. The problem with the human samples is that I have no control over how they are frozen! I have one sample from our path dept which was just put directly in the -80oC freezer the other sample I have is from LN2 storage (I presume it was snap frozen in LN2 after removal).

Anyway the sections from both samples are awful! They seem to cut ok but if you do any staining on them they fall apart and any tissue left just doesn't look right - the nuclei look misshapen, fuzzy, stretched and extracted. In sections from the tissue stored in LN2 the nuclei were either missing or didn't take up the dapi at all (except at the very edge of the section).

Has anyone had similar problems?

I cut 10um sections, air dry overnight, fix 100% acetone 10min.

Thanks
Sonya


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