Performance of fluorescence correlation spectroscopy for measuring diffusion and concentration

被引：159

作者：

Enderlein, J ^{[1
]}

Gregor, I ^{[1
]}

Patra, D ^{[1
]}

Dertinger, T ^{[1
]}

Kaupp, UB ^{[1
]}

机构：

[1] Forschungszentrum Julich, Inst Biol Informat Proc 1, D-52425 Julich, Germany

来源：

CHEMPHYSCHEM | 2005年 / 6卷 / 11期

关键词：

confocal microscopy; fluorescence spectroscopy; laser spectroscopy; photophysics; single-molecule spectroscopy;

D O I：

10.1002/cphc.200500414

中图分类号：

O64 [物理化学（理论化学）、化学物理学];

学科分类号：

070304 ; 081704 ;

摘要：

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.

引用

页码：2324 / 2336

页数：13

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