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Proceedings Paper

Imaging of rotational Brownian motion of biomolecules in living cells using fluorescence depolarization microscopy
Author(s): Shuji Toyonaga; Yoshitaro Nakano
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Paper 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.

Paper Details

Date Published: 29 April 1998
PDF: 12 pages
Proc. SPIE 3260, Optical Investigations of Cells In Vitro and In Vivo, (29 April 1998); doi: 10.1117/12.307085
Show Author Affiliations
Shuji Toyonaga, Lab. of Molecular Biophotonics (Japan)
Yoshitaro Nakano, Lab. of Molecular Biophotonics (Japan)

Published in SPIE Proceedings Vol. 3260:
Optical Investigations of Cells In Vitro and In Vivo
Robert C. Leif; Daniel L. Farkas; Robert C. Leif; Bruce J. Tromberg, Editor(s)

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