[Histonet] Manual fixation/processing of reconstructed skin cultures grown on polycarbonate filters

From:"Colin Smith"

Can anyone help?
I am currently setting up a basic histology system for analysing
reconstructed tissues grown in culture on polycarbonate filters, similar to
Mattek's Epiderm. We currently have begged borrowed or stolen,
two water baths (ambient - 100C), a circulating warm air oven, a rocking
microtome and cold blocks, along with glass staining jars/racks, forceps etc.
At the moment, equipment such as a wax dispenser, purpose built cooling
plates etc are out of the question until I can show that we can show that the
model we are growing is validated, which will include immunohistochemistry of
cytokeratin and cell adhesion markers etc. Thus we are in the catch 22
situation of having to prove something without the equipment needed to do the
job, in order to justify buying the equipment!! But thats accountants for
you, go figure.
Anyway, can anyone give any useful pointers/protocols for
fixing/processing/embedding/sectioning such tissues using such primative
Any help, no matter how trivial will be greatfully received.
Many Thanks in Advance,
Colin Smith BSc CBiol MIBiol Eurotox Registered Toxicologist
In Vitro Toxicologist 
Thor Specialities UK Ltd
Wincham Avenue,
Tel: +44 1606 818873
Fax: +44 1606 818801

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