JB4 aka GMA plastic staining

From:Gayle Callis <uvsgc@msu.oscs.montana.edu>

You can use most stains performed on paraffin sections, with the exception
of some trichromes, they tend to be more difficult.  However, there is a
modification of this stain for 1 um sections that work beautifully.

You can do Jones Methenamine silver, Giemsa, H&E, etc.  One thing that is
nice the plastic does not (cannot!) be removed, and excessive dehydration
in alcohols, clearing in xylene will cause plastic to crack.  We usually
air dried after staining, and coverslipped a DRY section with routine
mounting media.  

Eosin Phloxine counterstain is wonderful, can buy this from Richard Allan
or make up yourself.

Immunostaining is more difficult often not possible since plastic tend to
prevent immunoglobulins (hydrophobic problem) access to antigenic epitopes.

Enzyme histochemistry can also be performed, plus some technics for
embedding and lipid staining.

Take care, this stuff is toxic, work in a hood, with nitrile gloves, wear
safety glasses when sectioning (heard of a section flying into technicians
eye) keep skin contact down to very minimum, never with wet hands.  Very
sensitizing material, plus carcinogens in accelerator. 

There are publications on GMA in Journal of Histotechnology, back in 70's
Clyde Lulham publication and Nate Brinn on enzyme histochemistry, plus
others over the years.  

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

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