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Journal of Biomedical Optics

Dissecting eukaryotic cells by coherent phase microscopy: quantitative analysis of quiescent and activated T lymphocytes
Author(s): Vladimir P. Tychinsky; Aleksandr V. Kretushev; Tatiana V. Vyshenskaia; Alexander A. Shtil
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Paper Abstract

We present a concept for quantitative characterization of a functional state of an individual eukaryotic cell based on interference imaging. The informative parameters of the phase images of quiescent and mitogen-activated T lymphocytes included the phase thickness, phase volume, the area, and the size of organelles. These parameters were obtained without a special hypothesis about cell structure. Combinations of these parameters generated a "phase portrait" of the cell. A simplified spherical multilayer optic model of a T lymphocyte was used to calculate the refractivity profile, to identify structural elements of the image with the organelles, and to interpret the parameters of the phase portrait. The values of phase image parameters underwent characteristic changes in the course of mitogenic stimulation of T cells; thereby, the functional state of individual cells can be described using these parameters. Because the values of the components of the phase portrait are measured in absolute units, it is possible to compare the parameters of images obtained with different interference microscopes. Thus, the analysis of phase portraits provides a new and perspective approach for quantitative, real-time analysis of subcellular structure and physiologic state of an individual cell.

Paper Details

Date Published: 17 July 2012
PDF: 9 pages
J. Biomed. Opt. 17(7) 076020 doi: 10.1117/1.JBO.17.7.076020
Published in: Journal of Biomedical Optics Volume 17, Issue 7
Show Author Affiliations
Vladimir P. Tychinsky, Moscow State Institute of Radiotechnics, Electronics and Automation (Russian Federation)
Aleksandr V. Kretushev, Moscow State Institute of Radiotechnics, Electronics and Automation (Russian Federation)
Tatiana V. Vyshenskaia, Lomonosov Moscow State Univ. (Russian Federation)
Alexander A. Shtil, N. N. Blokhin Russian Cancer Research Ctr. (Russian Federation)


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