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Proceedings Paper

Multiparameter single molecule spectroscopy gives insight into the complex photophysics of fluorescence energy transfer (FRET) coupled biosystems
Author(s): F. Schleifenbaum; C. Blum; K. Elgass; S. Peter; V. Subramaniam; A. J. Meixner
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Paper Abstract

Since the discovery of the technique in the early 1990s, single molecule spectroscopy has been used as a powerful tool to investigate and characterize fluorescent molecules, revealing insights into molecular behavior far beyond the information content that can be obtained by conventional ensemble studies. Several spectroscopic techniques have been established at the single molecule level, including spectrally resolved fluorescence, fluorescence lifetime investigations, or single molecule Raman measurements. However, the combination of two or more of these spectroscopies applied to the same individual molecule in multiparameter approaches yields a deeper understanding of molecular systems. In this contribution, we present our results of combined spectrally- and time-resolved fluorescence microscopy of the intrinsic fluorescence energy transfer (FRET) system of the red fluorescent protein DsRed. Correlating the results obtained from the two spectroscopic techniques, we are able to determine all relevant parameters to describe the energy transfer processes within the DsRed system without any further assumptions. We further discuss fluorescence and surface enhanced Raman scattering (SERS) spectroscopy of the same individual DsRed unit, which can help to propose mechanisms for photodegeneration of the distinct chromophores involved.

Paper Details

Date Published: 20 February 2009
PDF: 8 pages
Proc. SPIE 7190, Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications, 71900U (20 February 2009); doi: 10.1117/12.809298
Show Author Affiliations
F. Schleifenbaum, Univ. of Tuebingen (Germany)
C. Blum, Univ. of Twente (Netherlands)
K. Elgass, Univ. of Tuebingen (Germany)
S. Peter, Univ. of Tuebingen (Germany)
V. Subramaniam, Univ. of Twente (Netherlands)
A. J. Meixner, Univ. of Tuebingen (Germany)

Published in SPIE Proceedings Vol. 7190:
Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications
Samuel Achilefu; Ramesh Raghavachari, Editor(s)

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