Jump to main content
Jump to site search

Issue 14, 2009
Previous Article Next Article

Manipulation of the local density of photonic states to elucidate fluorescent protein emission rates

Author affiliations

Abstract

We present experiments to determine the quantum efficiency and emission oscillator strength of exclusively the emitting states of the widely used enhanced green fluorescent protein (EGFP). We positioned the emitters at precisely defined distances from a mirror to control the local density of optical states, resulting in characteristic changes in the fluorescence decay rate that we monitored by fluorescence lifetime microscopy. To the best of our knowledge, this is the first emission lifetime control of a biological emitter. From the oscillation of the observed emission lifetimes as a function of the emitter to mirror distance, we determined the radiative and nonradiative decay rates of the fluorophore. Since only the emitting species contribute to the change in emission lifetimes, the rates determined characterize specifically the quantum efficiency and oscillator strength of the on-states of the emitter, in contrast to other methods that do not differentiate between emitting and dark states. The method reported is especially interesting for photophysically complex systems like fluorescent proteins, where a range of emitting and dark forms has been observed. We have validated the analysis method using Rhodamine 6G dye, obtaining results in very good agreement with the literature. For EGFP we determine the quantum efficiency of the on-states to be 72%. As expected for this complex system, our value is higher than that determined by methods that average over on- and off-states.

Graphical abstract: Manipulation of the local density of photonic states to elucidate fluorescent protein emission rates

Back to tab navigation

Publication details

The article was received on 13 Oct 2008, accepted on 12 Jan 2009 and first published on 11 Feb 2009


Article type: Paper
DOI: 10.1039/B817902F
Citation: Phys. Chem. Chem. Phys., 2009,11, 2525-2531
  •   Request permissions

    Manipulation of the local density of photonic states to elucidate fluorescent protein emission rates

    Y. Cesa, C. Blum, J. M. van den Broek, A. P. Mosk, W. L. Vos and V. Subramaniam, Phys. Chem. Chem. Phys., 2009, 11, 2525
    DOI: 10.1039/B817902F

Search articles by author

Spotlight

Advertisements