[Histonet] RE: Histonet Digest, Vol 128, Issue 14

[Joana Moreira]

James, Hi!
I used to use EDTA decalcification back in the UK and didn't deal with complaints from hematopathologists regarding that particular problem. The hospital where I worked didn't have the Milestone Bonestation but did use a temperature controlled oscillator performing the decalcification at 37C with oscillation for 48h. It's true it takes a lot longer to decalcify but the quality of IHC, ISH/FISH as well as DNA/RNA yield is far better.
The decalcification process is published in a paper: Fluorescence in situ hybridisation analysis of bone marrow trephine biopsy specimens; an additional tool in the diagnostic armoury; that you can find at http://www.ncbi.nlm.nih.gov/pubmed/23038690.
Your institution much probably has access to it but if you're interested and can't get the full version email me and I shall get that to you.

Joana

Joana Moreira
Supervisor - Anatomical Pathology
Department of Pathology

Sidra Medical & Research Center
PO Box 26999 | Doha, Qatar
Direct Line  +974-4404-2036
jmoreira <@t> sidra.org | www.sidra.org



Message: 3
Date: Mon, 14 Jul 2014 13:37:53 -0500
From: "Vickroy, Jim" <Vickroy.Jim <@t> mhsil.com>
Subject: [Histonet] Bone Marrow Trephine Cases
To: "histonet <@t> lists.utsouthwestern.edu"
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Over the past several years we have had some questions by our hematopathologists regarding the quality of the trephine blocks.  The biggest concern has been that the sections seem to have  a fragmented appearance which we have determined in the past to be areas where the lipid cell borders break.   We have found that the artifact is diminished when we pick the sections up as quickly as we can which led us to believe that the cell borders break when they are left on the   warm water in the waterbath.  Some of the pathologists still believe the artifact is caused by the decalcification process.

I have reviewed protocols before and obviously found that labs use different decalcifying agents from a harsh HCL solution to a weaker formic acid solution.  We currently use Immunocal which I believe is a formic acid solution.   I have also been introduced to some new instrumentation available such as the Milestone Bone Station which uses a EDTA solution with some added heat and agitation.  I realize that EDTA  yields more DNA and RNA which is obviously an advantage however the length of time for the decal process with EDTA is much longer.   I'm not sure that using EDTA for decalcification will however reduce the artifact that the pathologists see (fragmentation and breaking up of the lipid cells in the sections.

I would appreciate any information that others might be able to share from their experiences and if they have dealt with a similar problem.

Thanks



James Vickroy BS, HT(ASCP)

Surgical  and Autopsy Pathology Technical Supervisor
Memorial Medical Center
217-788-4046
217-415-7505 Cell




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