Open AccessConfocal DNA cytometry: A contour‐based segmentation algorithm for automated three‐dimensional image segmentation
Author(s) -
Beliën Jeroen A.M.,
van Ginkel Hielke A.H.M.,
Tekola Paulos,
Ploeger Lennert S.,
Poulin Neal M.,
Baak Jan P.A.,
van Diest Paul J.
Publication year - 2002
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/cyto.10138
Subject(s) - segmentation , confocal , artificial intelligence , cytometry , flow cytometry , image segmentation , computer science , pattern recognition (psychology) , confocal microscopy , computer vision , biology , microbiology and biotechnology , physics , optics
Abstract Background Confocal laser scanning microscopy (CLSM) presents the opportunity to perform three‐dimensional (3D) DNA content measurements on intact cells in thick histological sections. So far, these measurements have been performed manually, which is quite time‐consuming. Methods In this study, an intuitive contour‐based segmentation algorithm for automatic 3D CLSM image cytometry of nuclei in thick histological sections is presented. To evaluate the segmentation algorithm, we measured the DNA content and volume of human liver and breast cancer nuclei in 3D CLSM images. Results A high percentage of nuclei could be segmented fully automatically (e.g., human liver, 92%). Comparison with (time‐consuming) interactive measurements on the same CLSM images showed that the results were well correlated (liver, r = 1.00; breast, r = 0.92). Conclusions Automatic 3D CLSM image cytometry enables measurement of volume and DNA content of large numbers of nuclei in thick histological sections within an acceptable time. This makes large‐scale studies feasible, whereby the advantages of CLSM can be exploited fully. The intuitive modular segmentation algorithm presented in this study detects and separates overlapping objects, also in two‐dimensional (2D) space. Therefore, this algorithm may also be suitable for other applications. Cytometry 49:12–21, 2002. © 2002 Wiley‐Liss, Inc.