sectioning araldite

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From:"BrosnanWatters, Gayle" <>
To:'histonet' <>
Content-Type:text/plain; charset="windows-1252"

	I am not really an em tech, either, but a psychologist who sections
mouse brains embedded in araldite.  I float my sections on the slide in a
little distilled water, and then I take them to a hot plate (I set mine
about at the number 2, what ever temp that is!), and the procedure then is
really specific - you have to get the water hot enough to flatten out the
tissue which is in araldite (as the other gentleman said, it is hard and
won't flatten until it is hot) BUT you can't let the water bubble at all or
it will form bubbles UNDER your tissue.  After the tissue is lying flat, I
carefully drain the water off the slide by tipping it slightly and holding a
tissue or something just touching the slide.  It then has to dry completely
before I stain it.  I should say that I section at 1 micron (not for EM,
just for light microscopy) but the lab I used to be in used the same
technique (approximately) for EM.  I stain my tissue with a mixture of equal
parts of toludene blue, azure II, and borax.  (I know, I know, Azure II has
toludene blue in it, but for some reason this recipe works and it was the
one I was taught!) In order to stain, after the slide is dry, I reheat it
(it has to be quite hot, as this is what makes the tissue adhere to the
slide), then put the stain on the tissue, continue to heat just until it is
ALMOST ready to boil, but NOT boiling, then rinse off the excess stain.
This works for mouse and rat brains, but couldn't tell you if it is anything
you'd want.  Hey, I'm just a psychologist, what do I know???
	I am a new lister, and I LOVE this!!

Gayle Brosnan-Watters, Ph.D.
Assistant Professor
Psychology Department
Vanguard University of Southern California
55 Fair Drive
Costa Mesa, CA  92626
Phone 714-556-3610 Ext. 454
Fax 714-966-6316

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