DNA ploidy analysis in breast carcinoma. Comparison of unfixed and fixed tissue analyzed by image and flow cytometry.

Author(s): Ottesen GL, Christensen IJ, Larsen JK, Larsen J, Christiansen J, Baldetorp B, Linden T, Hansen B, Andersen JA

Publication: Anal Quant Cytol Histol, 1997, Vol. 19, Page 413-22

PubMed ID: 9349901 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of preservation method, slide type, and analysis by FCM versus ICM on DNA ploidy determination in breast carcinoma specimens.

   Summary of Findings:

   FCM of frozen tissue gave the highest resolution in DNA analysis with a median coefficient of variation (CV) of the DNA diploid G1 peak of 1.7%. Median CVs of the DNA diploid G1 peaks for FCM of formalin-fixed tissue and ICM of imprints from frozen specimens were 2.5% and 3.5%, respectively. ICM histograms of formalin-fixed tissue sections included poorly defined peaks, but of the defined ones, the CV was >10%. FCM of frozen tissue showed the highest number of DNA nondiploid clones compared to the other three methods, however, concordance was high between FCM of frozen tissue and each of the other three methods with respect to classification as diploid or nondiploid. ICM of imprints had the advantage of identifying clones with DNA indices of >3, and ICM, unlike FCM, could identify combined DNA diploid and nondiploid patterns and allowed for the analysis of small lesions. The authors state that use of 4',6-diamidino-2-phenylindole (DAPI) staining rather than propidium iodide for FCM of formalin-fixed tissue led to a higher frequency of recovery of non-diploid clones.

   Biospecimens

   • Tissue - Breast

   Preservative Types

   • Frozen
   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   DNA	Flow cytometry
   DNA	Light microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) Frozen
   Biospecimen Aliquots and Components	Type of slide	Imprint Tissue section
   Flow cytometry Specific	Technology platform	Light microscopy
   Flow cytometry Specific	Type of tissue stain	Propidium iodide DAPI

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