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Updated: Jun 16, 2026

Time-Resolved Fluorescence Anisotropy from Single Molecules for Characterizing Local Flexibility in Biomolecules
Published on: April 25, 2025
Statistical analysis of time resolved single molecule fluorescence data without time binning
1Institut für Physikalische Chemie, Universität Mainz, D-55099 Mainz, Germany. hinze@uni-mainz.de
Abstract:
We depict two algorithms to calculate correlation functions from two different time resolved single molecule fluorescence experiments without the need of time binning. Our first procedure allows to calculate the reduced linear dichroism from polarization resolved fluorescence data. Since we process single photon counts instead of time binned data, considerably faster fluctuations of the dichroism can be analyzed than with conventional methods. With our second procedure time resolved fluorescence obtained with a time correlated single photon counting setup can be analyzed with respect to fluorescence lifetime fluctuations. Again this new algorithm processes single photon events making time binning of photon counts obsolete. Both methods presented are characterized by enhanced time resolution thus allowing to study fast fluctuations of either single molecular orientation or fluorescence life times, respectively.

