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Proceedings Paper

Ab initio modeling of fluorescence fluctuation spectroscopy
Author(s): Joerg Enderlein; Martin Bohmer
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Paper 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.

Paper Details

Date Published: 19 June 2003
PDF: 11 pages
Proc. SPIE 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues, (19 June 2003);
Show Author Affiliations
Joerg Enderlein, Forschungszentrum Julich (Germany)
Martin Bohmer, Forschungszentrum Julich (Germany)

Published in SPIE Proceedings Vol. 4962:
Manipulation and Analysis of Biomolecules, Cells, and Tissues
Dan V. Nicolau; Joerg Enderlein; Robert C. Leif; Daniel L. Farkas, Editor(s)

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