Systematic evaluation of fluorescence correlation spectroscopy data analysis on the nanosecond time scale

Katrin Steger; Stefan Bollmann; Frank Noé; Sören Doose

doi:10.1039/C3CP50644D

Systematic evaluation of fluorescence correlation spectroscopy data analysis on the nanosecond time scale†

Katrin Steger,^a Stefan Bollmann,^a Frank Noé^b and Sören Doose*^a

* Corresponding authors

^a Department of Biotechnology and Biophysics, Biocenter, Am Hubland, Julius-Maximilians University, 97074 Würzburg, Germany
E-mail: soeren.doose@uni-wuerzburg.de
Fax: +49 931 3184509

^b Research Center Matheon, FU Berlin, Arnimallee 6, 14159 Berlin, Germany

Abstract

Signal fluctuations in a fluorescence time trace on nanosecond time scales can be induced by specific quenching interactions that report on the dynamics of biomolecules. Fluorescence correlation spectroscopy is an analysis tool to investigate dynamic processes on time scales from pico- to milliseconds or longer. Under certain conditions, e.g. in a solvent of high viscosity, a fluorescence labeled dynamic biomolecule yields multiple independent correlation decays due to rotational and translational diffusion, fluorescence quenching interactions, and fluorophore photophysics. We compared parameter estimation for FCS data with multiple correlation decays by dynamical fingerprint analysis and by the non-linear Levenberg–Marquardt fitting procedure and identified conditions for which dynamical fingerprint analysis can be of advantage. In this context we identified a previously unrecognized photophysical process in ATTO655 that introduces fluorescence intermittency on nanosecond time scales that is absent in MR121. The optimized fitting procedure is used to resolve the viscosity dependence of fluorescence quenching for photoinduced electron transfer probes.

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DOI: https://doi.org/10.1039/C3CP50644D
Article type: Paper
Submitted: 12 Feb 2013
Accepted: 26 Apr 2013
First published: 30 Apr 2013

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Phys. Chem. Chem. Phys., 2013,15, 10435-10445

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Systematic evaluation of fluorescence correlation spectroscopy data analysis on the nanosecond time scale

K. Steger, S. Bollmann, F. Noé and S. Doose, Phys. Chem. Chem. Phys., 2013, 15, 10435 DOI: 10.1039/C3CP50644D

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Physical Chemistry Chemical Physics

Systematic evaluation of fluorescence correlation spectroscopy data analysis on the nanosecond time scale†

Abstract

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Systematic evaluation of fluorescence correlation spectroscopy data analysis on the nanosecond time scale

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