Re: Tripp and MacKay
|From:||Gayle Callis <firstname.lastname@example.org>|
No, prefer to do a toludine blue stain or T blue combined with inhouse
prepared MacNeal tetrachrome on acid etched undecalcified bone sections
embedded in PMMA. OR a von Kossa counterstained with van Gieson.
You should be able to see osteiod quite well with an high quality H&E
decalcified section, using eosin phloxine instead of just eosin.
E/Phloxine always gave us better contrast in bone. A good toluidine blue
should also work.
Samples can only be 1 - 2mm thick, and 2% silver nitrate 48 hours, in dark.
There are caveats with Tripp Mackay - and having tried it once, I preferred
other staining technics. It gave us poor results even after following
You can overimpregnate silver at surface of bone and underimpregnate deeper
in sample. If you recognize these problems, you can deal with it, probably
can't section deep into the sample or do serial sections.
Don't use nitric or HCl acids to decalcify, 10% formic is recommended.
Strong acids can ruin silver deposits.
If you need to demonstrate cellular detail, do an unimpregnated, adjacent
I think 95% alcohol as a fixative is harsh on bone, 70% ethanol has long
been used as a bone fixative, but it makes morphology less than optimal,
dessicates the soft tissues/cells, and 95% would be worse. Fixing in 70%
would help rehydrate since it takes many distilled water changes at 4
hours/change before staining begins.
NBF fixed bone could be used, but you have to insure ALL formalin is
removed, using distilled water, before you can attempt impregnation. How
long you rinse is a good question, could be many changes for hours at a
time to insure? total removal of formalin.
At 12:32 PM 5/17/01 +0000, you wrote:
>Does anyone out there use the Tripp and MacKay block impregnation method for
>osteoid seams? If so how long do you leave it in the dark in the silver
>solution? Does anyone use an alternative?
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Veterinary Molecular Biology
Montana State University - Bozeman
Bozeman MT 59717-3610
404 994-4303 (FAX)
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