Nicola,
        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.
                                       Regards,
                                                    Tony
Histopathology Group
Asthma Biology Department.
RIRP CEDD.
GlaxoSmithKline Medicines Research Centre,
Gunnelswood Road,
STEVENAGE,
Hertfordshire.
  SG1 2NY
tel.          +44 (0)1438 764117
fax.         +44 (0)1438 764782
email.    Tony.J.Savage@gsk.com
mobile    +44 07753609835





ncragg 

24-Mar-2003 11:58

 
 

        To:     "'HistoNet Server'"

        cc: 
        Subject:        Processing cell monolayer/ multi-layer cultures

Can anyone offer any advice on processing cell cultures, grown in 
monolayer 
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 
cell 
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.

Thank you

Nicola Cragg
Epistem Ltd,
Manchester, UK