rat vetebrae/Ehrlich

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu> (by way of histonet)
To:histonet <histonet@magicnet.net>
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I tried Ehrlichs hematoxylin on decalcified bone, with excellent
results.  One of the joys of using this hematoxylin is NO MERCURY
in the recipe.  I ripened the Ehrlich rapidly with sodium iodate,
instead of weeks exposure to the air.  This hematoxylin gave some of my
best stained bone sections ever and when compared with the Harris,
my pathologist actually liked it better!

Harris's also works, best when fresh.  You can
also do an eosin counterstain, shorten time a bit to not overstain
the bone.  I stained for 10 minutes in both of these hematoxylins
to help counteract the effects of acid decalcification on the nuclei.

You can sometimes eliminate the differentiation step, just stain then
blue, to increase some tinctoral qualities in the cartilage.
I had poor results with commercially prepared Harris's, made my own and
loved the one that had mercury in in, alas, no more.  You can also
use hematoxylin 1 from Richard Allan, and if needed just stain
longer, a favored method.  They will send sample of all their
hematoxlins in a sample kit, always worth a try.

Just avoid overexposure (aka overdecalcification!) of acid on the
bone, preferrably with a decalcification endpoint check.

Good luck,
Gayle Callis









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