[Histonet] Fixation and Decalcification of Bone Marrow Bxs.

[Katri Tuomala]

Hi Charlene,
I think, if you fixed first with the 10% NBF properly and THEN decalcified
with EDTA, your morphology would be great and EDTA decalcification does not
affect staining or imuunohistochemistry. The reason people do these things
together is to cut down the time, but you always pay for it. In this case
with poor morphology.
Cheers!

Katri

Katti Tuomala
Dept. of Pathology
St.Josep's Health Care
Hamilton, Ontario, Canada

----- Original Message ----- 
From: "Henry, Charlene" <Charlene.Henry <@t> STJUDE.ORG>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Friday, May 14, 2004 4:34 PM
Subject: [Histonet] Fixation and Decalcification of Bone Marrow Bxs.


Fellow Histotechs,
I would like to get some feedback on fixatives and decalcification
protocols used in your laboratories. Our pathologists, like many, want
perfection; so here is my problem. We were fixing our bone marrow bxs.
in 10% NBF and then decalcification for 1 hour prior to processing.
Because of the facility I work at, we perform IHC and ISH on a large
percentage of our bxs. With the method we were using, some of our
antibodies would not work on tissue that had been decalcified and the
decalcification completely destroyed the RNA making it impossible to
perform ISH. About 6 months ago, we switched to fixing our bone marrows
in 10% NBF saturated with EDTA (this fixes and decals at the same time)
with beautiful results with our IHC and also ISH. Even though our
pathologists are elated that the IHC and ISH are great, they are not
completely happy with the morphology of the H&E slides. I was just
wondering if anyone has any suggestions so we can accomplish both great
H&E morphology and great stains with IHC and ISH.
Thanks,

Charlene Henry, HT QIHC
Histology/IHC Section Head
Department of Pathology
St. Jude Children's Research Hospital
332 North Lauderdale
Memphis, TN  38105
Phone: 901-495-3349
FAX: 901-495-3100


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