Multiparameter Single Molecule Spectroscopy gives insight into the complex Photophysics of Fluorescence Energy Transfer (FRET) coupled Biosystems
| Autor: | Schleifenbaum, F., Blum, Christian, Elgass, K., Peter, S., Subramaniam, Vinod, Meixner, A.J., Achilefu, Samuel, Raghavachari, Ramesh |
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| Přispěvatelé: | Faculty of Science and Technology, Nanobiophysics |
| Rok vydání: | 2009 |
| Předmět: |
Chemistry
Single molecule SERS METIS-260255 Nanotechnology Surface-enhanced Raman spectroscopy Fluorescence in the life sciences IR-78009 Fluorescence spectroscopy symbols.namesake Förster resonance energy transfer Resonance fluorescence Chemical physics symbols FRET autofluorescent protein DsRed Fluorescence cross-correlation spectroscopy Raman spectroscopy Laser-induced fluorescence |
| Zdroj: | Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications, 719000-71901 STARTPAGE=719000;ENDPAGE=71901;TITLE=Reporters, Markers, Dyes, Nanoparticles, and Molecular Probes for Biomedical Applications |
| Popis: | 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 |
| Databáze: | OpenAIRE |
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