CONFOCAL 3-DIMENSIONAL DNA IMAGE CYTOMETRY THICK TISSUE-SECTIONS

We present a three-dimensional confocal DNA image cytometry (3-D CICM) method for analysis of DNA content in 30-40-mu m-thick sections of ro utinely processed paraffin-embedded specimens. A comparison of DNA plo idy 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 tiss ue sections used in conventional ICM, which suggested that many nuclei measured by this technique may be incomplete, This artifact was exclu ded in 3-D CICM by automatic rejection of cut nuclear profiles, This a nd the favorable ratio of tissue thickness to nuclear size in 3-D CICM permitted the DNA quantitation even in large cells with highly increa sed DNA ploidy values such as megakaryocytes and Reed-Sternberg cells of Hodgkin's disease. Additionally, 3D-CICM allowed evaluation of morp hometric parameters and S-dimensional reconstruction of studied cells. (C) 1996 Wiley-Liss, Inc.