Effects of Refractive Index and Viscosity on Fluorescence and Anisotropy Decays of Enhanced Cyan and Yellow Fluorescent Proteins

@article{Borst2005EffectsOR,
  title={Effects of Refractive Index and Viscosity on Fluorescence and Anisotropy Decays of Enhanced Cyan and Yellow Fluorescent Proteins},
  author={Jan Willem Borst and Mark A. Hink and Arie van Hoek and Antonie J.W.G. Visser},
  journal={Journal of Fluorescence},
  year={2005},
  volume={15},
  pages={153-160}
}
The fluorescence lifetime strongly depends on the immediate environment of the fluorophore. Time-resolved fluorescence measurements of the enhanced forms of ECFP and EYFP in water–glycerol mixtures were performed to quantify the effects of the refractive index and viscosity on the fluorescence lifetimes of these proteins. The experimental data show for ECFP and EYFP two fluorescence lifetime components: one short lifetime of about 1 ns and a longer lifetime of about 3.7 ns of ECFP and for EYFP… 

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References

SHOWING 1-10 OF 26 REFERENCES
The Influence of Solvent Viscosity on the Fluorescence Decay and Time-Resolved Anisotropy of Green Fluorescent Protein
This report describes fluorescence decay and time-resolved anisotropy studies of green fluorescent protein (GFP) in various environments. The addition of glucose and fructose, NaCl, or polyethylene
Effect of the Solvent Refractive Index on the Excited-State Lifetime of a Single Tryptophan Residue in a Protein
Excited-state lifetime and quantum yield of a fluorophore are determined by the radiative and nonradiative decay rates. There is an extensive literature about factors affecting the nonradiative decay
Principles of fluorescence spectroscopy
TLDR
This book describes the fundamental aspects of fluorescence, the biochemical applications of this methodology, and the instrumentation used in fluorescence spectroscopy.
An improved cyan fluorescent protein variant useful for FRET
TLDR
Cerulean is 2.5-fold brighter than ECFP and replacement of ECFP with Cerulean substantially improves the signal-to-noise ratio of a FRET-based sensor for glucokinase activation.
Fluorescence dynamics of green fluorescent protein in AOT reversed micelles.
Theory of fluorescence depolarization in macromolecules and membranes
A comprehensive formalism is developed to describe the decay of the fluorescence emission anisotropy r(t) in macroscopically isotropic systems where both excited state and orientational dynamics
Application of a reference convolution method to tryptophan fluorescence in proteins. A refined description of rotational dynamics.
TLDR
Fluorescence lifetime determinations for p-terphenyl, p-bis[2-(5-phenyloxazolyl)]benzene and N-acetyltryptophanamide show that with this method more reliable fits of the decays can be made than with the scatterer method, which is most frequently used.
Imaging the environment of green fluorescent protein.
...
1
2
3
...