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Analytical Cellular Pathology
Volume 14 (1997), Issue 1, Pages 41-49
http://dx.doi.org/10.1155/1997/539864

DNA Measurement of Overlapping Cell Nuclei in Thick Tissue Sections

Liang Ji1 and James Tucker2

1MRC Human Genetics Unit, Crewe Road, Edinburgh, EH4 2XU, UK
2Edinburgh University Department of Pathology, Teviot Place, Edinburgh EH8 9AG, UK

Received 7 August 1995; Revised 15 February 1997; Accepted 19 March 1997

Copyright © 1997 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The paper describes an improved image analysis procedure for measuring the DNA content of cell nuclei in thick sections of liver tissue by absorption densitometry. Whereas previous methods only permitted the analysis of isolated nuclei, the new technique enables both isolated and overlapping nuclei to be measured. A 3D segmentation procedure determines whether each object is an isolated nucleus or a pair of overlapping nuclei; in the latter case the combined optical density is redistributed to the individual nuclei. A selection procedure ensures that only complete nuclei are measured.

The method has been tested on specially‐prepared Feulgen‐stained 20μ sections of normal liver tissue. The overall distribution of the nuclear DNA measurements shows well‐defined diploid and tetraploid peaks, with coefficient of variations of less than 10%. Similar distributions were obtained from both the isolated nuclei and overlapped nuclei sub‐populations.