Multiparameter Single Molecule Spectroscopy gives insight into the complex Photophysics of Fluorescence Energy Transfer (FRET) coupled Biosystems

F. Schleifenbaum, Christian Blum, K. Elgass, S. Peter, Vinod Subramaniam, A.J. Meixner

Research output: Chapter in Book/Report/Conference proceedingConference contributionAcademicpeer-review

3 Citations (Scopus)


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
Original languageUndefined
Title of host publicationReporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications
EditorsSamuel Achilefu, Ramesh Raghavachari
Place of PublicationSan Jose
ISBN (Print)9780819474360
Publication statusPublished - 12 Feb 2009
EventReporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications - San Jose, CA
Duration: 26 Jan 200926 Jan 2009

Publication series

NameProceedings of SPIE
ISSN (Print)1605-7422


ConferenceReporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications
Other26 Jan 2009


  • FRET
  • autofluorescent protein
  • METIS-260255
  • Single molecule
  • IR-78009
  • DsRed
  • SERS

Cite this