Imaging of rotational Brownian motion of biomolecules in living cells using fluorescence depolarization microscopy

Shuji Toyonaga; Yoshitaro Nakano

doi:10.1117/12.307085

29 April 1998 Imaging of rotational Brownian motion of biomolecules in living cells using fluorescence depolarization microscopy

Shuji Toyonaga, Yoshitaro Nakano

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Proceedings Volume 3260, Optical Investigations of Cells In Vitro and In Vivo; (1998) https://doi.org/10.1117/12.307085
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

Abstract

Two-dimensional fluorescence depolarization measurement was achieved by a fluorescence microscope equipped with polarizing devices and an image intensified CCD camera. Anisotropy images were acquired using living cells stained with a membrane specific binding dye. It was found that the bound dye is spatially distinguishable from the free dye which does not bind to the membrane of a cell, due to the difference in rotational Brownian motion. In addition, we succeeded in obtaining fluorescence depolarization images by means of time- resolved measurement and two-photon excitation measurement. Two-photon excitation images showed a superior signal-to-noise ratio compared to one-photon excitation images. Fluorescence depolarization imaging will therefore prove a powerful tool for studying molecular functions in cells.

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Shuji Toyonaga and Yoshitaro Nakano "Imaging of rotational Brownian motion of biomolecules in living cells using fluorescence depolarization microscopy", Proc. SPIE 3260, Optical Investigations of Cells In Vitro and In Vivo, (29 April 1998); https://doi.org/10.1117/12.307085

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KEYWORDS

Anisotropy

Luminescence

Signal to noise ratio

Molecules

Image quality

Microscopy

Fluorescence anisotropy

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