Abstract

In this paper we analyze factors influencing the accuracy and precision of optical density measurement of light microscopical objects. The study is applied to the DNA content of Feulgen-stained cells using a low-resolution, TV-equipped microscope connected to a digital image processing system. Factors influencing the accuracy of density measurement include staining (not considered here), image formation (glare, focus, diffraction), image sampling (distribution, sampling density, noise), and computational accuracy. These factors are reviewed with respect to potential remedies. Evaluation of the measurement error contribution is performed on the level of the individual cell and on the specimen level. It is concluded that a coefficient of variation of 5% (in contrast to 3.5% using flow cytometry of the same specimen) in the measured values of normal cells of one specimen is attainable using adequate shading correction. With relatively simple computational methods, image cytometry may be well suited for pathology practice.

© 1987 Optical Society of America

Automated densitometry of cell populations in a continuous-motion imaging cell scanner

James H. Tucker, O. A. N. Husain, Keith Watts, Stephen Farrow, Royston Bayley, and Margaret H. Stark
Appl. Opt. 26(16) 3315-3324 (1987)

Image DNA-index (ploidy) analysis in cancer diagnosis

P. S. Oud, A. G. J. M. Hanselaar, M. M. M. Pahlplatz, J. W. R. Meijer, and G. Peter Vooijs
Appl. Opt. 26(16) 3349-3355 (1987)

Optical microscope system for standardized cell measurements and analyses

James W. Bacus and Les J. Grace
Appl. Opt. 26(16) 3280-3293 (1987)

High performance pathology workstation using an automated multispectral microscope

Harvey L. Kasdan, Kenneth C. Langford, John Liberty, Michael Zachariash, and Fred H. Deindoerfer
Appl. Opt. 26(16) 3294-3300 (1987)

Computer-aided microscopy in medicine: introduction to the 15 August 1987 issue of Applied Optics

Norman J. Pressman
Appl. Opt. 26(16) 3199-3209 (1987)