AW: [Histonet] FISH on B.M.

[Gudrun Lang]

Miriam,
do you do FISH on bone marrow trephine biopsies, that have to be decalcified
before cutting and staining?
If decalcifing is performed with acid like formic acid or even hydrochloric
acid, DNA is degraded to smaller fragments. Successfull hybridization of
this fragments is kind of chance, because it could easily happen that the
wanted DNA-sequence is destroyed. If the DNA-sequence is destroyed, there is
no way to restore it. Decalcifing should be performed with EDTA for a good
preservation of DNA. 

I have performed CISH on formic acid - decalcified bone marrow trephines
with success. But we demonstrated mRNA of light chains, that are usually
found in a big amount in the cytoplasm. So degrading a part of the mRNA
would not influence the final result. Our protocol for BMTB is one day
fixation in NBF, next day decal with 5-10% formic acid with 5-10%
formaldehyde for 8 hours, then processing like the other routine specimens.

Hope this helps
Gudrun Lang

Histolab, AKH Linz, Austria



-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von
birnbaumm <@t> asaf.health.gov.il
Gesendet: Sonntag, 01. November 2009 10:50
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] FISH on B.M.

Helo all,

 

We do FISH assay on FFPE tissue with "spot light tissue pretreatment" of
invitrogen. It do works on most of the tissues, but not on Bone Marrow
tissue. 

Do you have any idea?

 

Thanks

 

Dr. Miriam Birnbaum

Pathology 

Asaf Hrofeh Medical Center

Israel

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