[Histonet] Cell block preparation (from cultured cells)

[Jonathan Cremer]

Hi everyone,

Someone in the lab wishes to look at cultured cells in paraffin embedded cell blocks. These cells grow mostly in non-adherent clumps or cell bodies, which need to be preserved. Sections will be stained by IHC.
I've been looking into the matter, and it seems there is not really a single straightforward method for this (as is always the case in histoland, apparently).

I've been thinking to try fixing the cells in either 10% alcoholic formalin for 12-24 hours, or zinc-formalin. Then embed in agarose, fix some more with buffered formaldehyde and process.
- What are your thoughts on the fixative of choice?
- Should I process slowly, manually; or is a 'standard' ON processor protocol adequate (I don't know the exact timings, but I think it's a twelve hour protocol on a caroussel type processor).
- Would Histogel have any added benefit over agarose?

Thank you,
Jonathan

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Jonathan Cremer
Laboratory Technician
TARGID - KU Leuven