Re: Processing cell monolayer/ multi-layer cultures
I would strongly suggest that you consider using methacrylate
processing for cell monolayers and multilayer cultures. You get much
better cellular resolution than you do with paraffin wax. The important
thing to remember is to exclude the air and keep the temperature from
rising too much during polymerisation, otherwise the methacrylate goes
soft. I use to use a dessicator containing solid CO2 beneath a grid on which the polymerising resin blocks could be places.
The lid could then be put on the dessicator and the solid CO2 excluded the air.
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To: "'HistoNet Server'"
Subject: Processing cell monolayer/ multi-layer cultures
Can anyone offer any advice on processing cell cultures, grown in
or multi-layers with inserts / membranes? Are there any special
requirements? Would a general processing schedule used for animal (mainly
murine & rat) tissues be OK? Or would this be too rigorous, since the
layers are more delicate in nature than lumps of tissue? We use an
overnight schedule of 16 hours without vacuum. Is there such a thing as
over-processing? I read in the archives that overprocessing is a myth.
What are the symptoms that people claim are due to overprocessing? Any
help or advice would be much appreciated.
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