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Single-Molecule Surface-Enhanced Resonance Raman Spectroscopy of the Enhanced Green Fluorescent Protein EGFP

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Surface-Enhanced Raman Scattering

Part of the book series: Topics in Applied Physics ((TAP,volume 103))

4 Summary

We have investigated SM-SERRS spectra and intensity trajectories of individual EGFP molecules adsorbed on silver colloids. Spectral jumps, in a timescale of seconds, were observed in the SM-SERRS spectral trajectories. These jumps were attributed to a slow, reversible conversion between the protonated and deprotonated form of the chromophore in EGFP based on comparison with an ensemble Raman spectra of EGFP at different pH. The timescale of the conversion seems to match the timescale of the (long) off-periods observed in SM-fluorescence intensity trajectories. SM-SERRS intensity trajectories show, beside the slow fluctuations of the order of seconds, several faster timescales of fluctuations. We present a possible rational for those fluctuations based on literature data. We demonstrate that SM-SERRS data can be used to elucidate some of the complex photophysical processes occurring in GFPs.

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Author information

Authors and Affiliations

  1. Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200F, 3001, Heverlee, Belgium

    Satoshi Habuchi

  2. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, SGM209, Boston, MA, 02115, USA

    Johan Hofkens

Authors
  1. Satoshi Habuchi
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  2. Johan Hofkens
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Editor information

Editors and Affiliations

  1. Wellman Center for Photomedicine Harvard-M.I.T. Division of Health Sciences & Technology, Harvard University, Medical School, Boston, MA, 02114, USA

    Katrin Kneipp

  2. Department of Chemistry and Biochemistry, University of California Santa Barbara, Santa Barbara, CA, 93105, USA

    Martin Moskovits

  3. Wellman Center for Photomedicine, Harvard University, Medical School, Boston, MA, 02114, USA

    Harald Kneipp

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© 2006 Springer-Verlag Berlin Heidelberg

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Habuchi, S., Hofkens, J. (2006). Single-Molecule Surface-Enhanced Resonance Raman Spectroscopy of the Enhanced Green Fluorescent Protein EGFP. In: Kneipp, K., Moskovits, M., Kneipp, H. (eds) Surface-Enhanced Raman Scattering. Topics in Applied Physics, vol 103. Springer, Berlin, Heidelberg . https://doi.org/10.1007/3-540-33567-6_15

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