Open AccessThree‐dimensional DNA image cytometry by confocal scanning laser microscopy in thick tissue blocks of prostatic lesions
Author(s) -
Irinopoulou Theano,
Vassy Jany,
Beil Michael,
Nicolopoulou Polyxeni,
Encaoua David,
Rigaut Jean Paul
Publication year - 1997
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19970201)27:2<99::aid-cyto1>3.0.co;2-f
Subject(s) - cytometry , confocal , pathology , flow cytometry , confocal microscopy , biology , carcinoma , nuclear dna , hyperplasia , microbiology and biotechnology , medicine , optics , physics , mitochondrial dna , biochemistry , gene
DNA ploidy provides important information for the evaluation of the prognosis of prostate cancer. For the purpose of DNA cytometry and nuclear measurements, we developed an image processing system for the acquisition and processing of three‐dimensional (3D) images based on confocal scanning laser microscopy (CSLM). The advantage of the CSLM is the preservation of the tissue architecture and the possibility of multilabeling. It is possible to determine both individual nuclear features and cellular features and the degree of the spatial heterogeneity of several markers. Special attention was paid to the development of the automatic method for the 3D segmentation of cell nuclei. Thick tissue slides (100 μm), stained for DNA with chromomycin A 3 , from 4 patients (with benign hyperplasia, prostatic intraepithelial neoplasia (PIN), and well‐and poorly‐differentiated adenocarcinoma of the prostate), were studied in order to test the practicability of the developed methodology. DNA histograms showed a single peak in the diploid range for the hyperplasia and PIN cases. For the case of well‐differentiated carcinoma, 2 peaks were observed, 1 in the diploid range and 1 in the tetraploid range. The case of poorly‐differentiated carcinoma was characterized by an aneuploid distribution. For the cases of PIN and carcinomas, we observed a considerable variation of the volume of nuclei. Cytometry 27:99–105, 1997. © 1997 Wiley‐Liss, Inc.