Re: [Histonet] H&E staining on GMA plastic
After sectioning, merely dry the sections at 60C.
Gill 3 for 10 min up to 20 min. We never used Harris hematoxylin, only
progressive hematoxylins. Other progressive hematoxylins will work, i.e.
Richard Allan Hematoxylin 1. Bone requires more time in hematoxylin and
less time in eosin/phloxine, this counterstain gives wonderful contrast
with GMA sections.
Rinse 3 changes distilled water 10 dips each
Blue with Scotts tap water substitute 2 minutes
Rinse 3 changes distilled water 10 dips
AIR DRY SECTIONS!!!!
eosin/phloxine for 10 minutes, you may find this shorter time even down to
Rinse very quickly in 95% ethanol
Rinse very quickly in 100% ethanol
AIR DRY SECTIONS and mount coverslip WITHOUT dipping in xylene. If there
any funky wrinkles of plastic close to section, remove carefully with a
razor blade, sometimes we had curled lips on edges of sections - this way
you have a really flat coverglass.
Water and solvents (alcohol and xylenes, etc) soften GMA but do NOT remove
it, and will cause really ugly wrinkles and cracked plastic. We learned
to coverslip without wetting a slide with solvent.
The joy of GMA, hydrating sections is NOT necessary, you can immerse dry
sections directly into stains, hence the lovely short protocol.
By the way, I have some gems for GMA staining, a manual from an expert from
long ago, Sharon van de Velde back in the early 80's and also Nate Brinn -
he did some very fine work with this plastic, including enzyme
histochemical staining as did Sharon.
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)
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