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Proceedings Paper

Characterization of particle motions by fluorescence correlation spectroscopy with traveling interference fringe excitation
Author(s): Mineyuki Hattori; Hiroshi Yokoyama; Hideaki Shimizu
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Paper Abstract

A new improved fluorescence correlation spectroscopy method for determining motional states of particles, fluorescence correlation spectroscopy with traveling interference fringe excitation (FCSTFE), is presented. In this method the modulated fluorescence signal from particles excited by an interference fringe moving at constant velocity is detected, and cosine and sine Fourier coefficients at the frequency of the traveling fringe are recorded. Autocorrelation functions of Fourier coefficients and the square of amplitude were found to express the motion of the fluorescent particles and to correspond to that of laser light scattering experiment in homodyne mode. The apparatus utilizing a photon counting system was constructed and the performance of the system in measuring diffusion coefficients of fluorescent polystyrene spheres suspended in water and ethylene glycol mixed solvents was demonstrated. The FCSTFE experiments using biological membranes are expected to yield similar information to that obtained from ideal laser light scattering experiments which are difficult to perform because of the complexity in data analysis due to stray light and multiple scattered light from the samples. The new method is also superior in avoiding severe problems, such as the damage in samples caused by the intense bleaching light, which may occur during the fluorescence photobleaching recovery.

Paper Details

Date Published: 25 March 1996
PDF: 8 pages
Proc. SPIE 2705, Fluorescence Detection IV, (25 March 1996); doi: 10.1117/12.236178
Show Author Affiliations
Mineyuki Hattori, Electrotechnical Lab. (Japan)
Hiroshi Yokoyama, Electrotechnical Lab. (Japan)
Hideaki Shimizu, Electrotechnical Lab. (Japan)


Published in SPIE Proceedings Vol. 2705:
Fluorescence Detection IV
E. Roland Menzel; Abraham Katzir, Editor(s)

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