期刊
CHEMPHYSCHEM
卷 6, 期 11, 页码 2324-2336出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cphc.200500414
关键词
confocal microscopy; fluorescence spectroscopy; laser spectroscopy; photophysics; single-molecule spectroscopy
Fluorescence correlation spectroscopy (FCS) has become an important tool for measuring diffusion, concentration, and molecular interactions of cellular components. The interpretation of FCS data critically depends on the measurement set-up. Here, we present a rigorous theory of FCS based on exact wave-optical calculations. Six of the most important optical and photophysical factors that influence FCS are studied: fluorescence anisotropy, cover-slide thickness, refractive index of the sample, laser-beam geometry, optical saturation, and pinhole adjustment. Our theoretical framework represents a general attempt to link all relevant parameters of the experimental set-up with the measured correlation function.
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