Confocal 3-dimensional DNA image cytometry in thick tissue sections

Magdalena Czader, Anders Liljeborg, Gert Auer, Anna Porwit

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

We present a three-dimensional confocal DNA image cytometry (3-D CICM) method for analysis of DNA content in 30-40-μm-thick sections of routinely processed paraffin-embedded specimens. A comparison of DNA ploidy profiles obtained by 3-D CICM and conventional DNA image cytometry (ICM) on tissue sections showed significantly higher numbers of cells with high DNA content in DNA histograms by 3-D CICM. As estimated by 3-D CICM, the size of nuclei frequently exceeded the thickness of tissue sections used in conventional ICM, which suggested that many nuclei measured by this technique may be incomplete. This artifact was excluded in 3-D CICM by automatic rejection of cut nuclear profiles. This and the favorable ratio of tissue thickness to nuclear size in 3-D CICM permitted the DNA quantitation even in large cells with highly increased DNA ploidy values such as megakaryocytes and Reed-Sternberg cells of Hodgkin's disease. Additionally, 3D-CICM allowed evaluation of morphometric parameters and 3-dimensional reconstruction of studied cells.

Original languageEnglish (US)
Pages (from-to)246-253
Number of pages8
JournalCytometry
Volume25
Issue number3
DOIs
StatePublished - Nov 1 1996
Externally publishedYes

Keywords

  • 3-Dimensional reconstruction
  • Confocal microscopy
  • DNA image cytometry
  • DNA ploidy
  • Nuclear morphometry

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Biophysics
  • Hematology
  • Endocrinology
  • Cell Biology

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