Effect of bone decalcification procedures on DNA in situ hybridization and comparative genomic hybridization. EDTA is highly preferable to a routinely used acid decalcifier.

Author(s): Alers JC, Krijtenburg PJ, Vissers KJ, van Dekken H

Publication: J Histochem Cytochem, 1999, Vol. 47, Page 703-10

PubMed ID: 10219063 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to compare the effects of acid-based (RDO) and chelating-agent (EDTA) decalcification on ISH, CGH and flow cytometry results.

   Summary of Findings:

   While strong ISH signals were detected using centromereic probes on EDTA-decalcified specimens, RDO-decalcified specimens failed to show any signal. The authors report that there were too many nuclear debris to determine the DNA index by flow cytometry from RDO-decalcified specimens; however, EDTA-decalcified specimens showed distinct diploid and tetraploid peaks. DNA was not extracted from the RDO-treated bone, but fragments of up to 10 kB were extracted from the EDTA-decalcified specimens. Further, using EDTA-decalcified specimens, determination of loss and gain of sequences was possible by CGH.

   Biospecimens

   • Tissue - Bone

   Preservative Types

   • Formalin

   Diagnoses:

   • Autopsy
   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   DNA	Flow cytometry
   DNA	In situ hybridization
   DNA	CGH

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Decalcification solution/ duration	Acid based agent (RDO) Chelating agent (10% EDTA)

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