Research Papers: Imaging

Dissecting eukaryotic cells by coherent phase microscopy: quantitative analysis of quiescent and activated T lymphocytes

[+] Author Affiliations
Vladimir P. Tychinsky, Alexander V. Kretushev

Moscow Institute of Radioengineering, Electronics and Automation, Laboratory of Coherent Phase Microscopy, 78 Vernadsky Avenue, Moscow 119454, Russia

Tatiana V. Vyshenskaya

Moscow State University, Department of Physics, Vorob’evy Gory, Moscow 119992, Russia

Alexander A. Shtil

Blokhin Cancer Center, 24 Kashirskoye Shosse, Moscow 115478, Russia

J. Biomed. Opt. 17(7), 076020 (Jul 17, 2012). doi:10.1117/1.JBO.17.7.076020
History: Received March 27, 2012; Revised May 23, 2012; Accepted June 18, 2012
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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.

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© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

Vladimir P. Tychinsky ; Alexander V. Kretushev ; Tatiana V. Vyshenskaya and Alexander A. Shtil
"Dissecting eukaryotic cells by coherent phase microscopy: quantitative analysis of quiescent and activated T lymphocytes", J. Biomed. Opt. 17(7), 076020 (Jul 17, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.7.076020


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