Ab initio modeling of fluorescence fluctuation spectroscopy

Joerg Enderlein; Martin Bohmer

doi:10.1117/12.478906

19 June 2003 Ab initio modeling of fluorescence fluctuation spectroscopy

Joerg Enderlein, Martin Bohmer

Proceedings Volume 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues; (2003) https://doi.org/10.1117/12.478906
Event: Biomedical Optics, 2003, San Jose, CA, United States

Abstract

Fluorescence correlation spectroscopy (FCS) has developed into a widely used and very successful spectroscopic technique for detecting and quantifying minute concentrations of fluorescing dyes in solution, as well as for obtaining information about molecular parameters such as diffusion coefficients, photophysical transition rates, or chemical reaction kinetics. The standard evaluation method of FCS data is based on the simplified assumption that the convolution of the excitation intensity distribution and the light collection efficiency function is a three-dimensional Gaussian distribution. Although that assumption leads to satisfactory fits of experimental data, the adjustable parameters of this standard model have a rather unphysical meaning. In the present paper, an ab initio approach to modeling FCS curves is developed that is based on exact wave-optical calculations of fluorescence excitation and detection. Comparison with the standard model is made, and the ab initio calculated model curves are used for fitting experimental data and deriving absolute values of diffusion coefficient and concentration.

Citation Download Citation

Joerg Enderlein and Martin Bohmer "Ab initio modeling of fluorescence fluctuation spectroscopy", Proc. SPIE 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues, (19 June 2003); https://doi.org/10.1117/12.478906

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