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Related Experiment Video

Updated: Feb 18, 2026

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

Published on: February 9, 2012

Quantifying Microsecond Transition Times Using Fluorescence Lifetime Correlation Spectroscopy.

Arindam Ghosh¹, Sebastian Isbaner¹, Manoel Veiga-Gutiérrez²

¹III. Institute of Physics, Georg August University , 37077 Göttingen, Germany.

The Journal of Physical Chemistry Letters

|November 30, 2017

Summary

This summary is machine-generated.

Fluorescence lifetime correlation spectroscopy (FLCS) reveals microsecond dynamics in enhanced green fluorescent protein (EGFP). This method tracks rapid transitions between fluorescent states, crucial for understanding complex light-emitting systems.

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Area of Science:

Photophysics
Biophysics
Spectroscopy

Background:

Complex luminescent emitters, like fluorescent proteins, often possess multiple emitting states.
These states cause rapid fluctuations in excited-state lifetime, complicating analysis.
Enhanced green fluorescent protein (EGFP) is a widely used model system.

Purpose of the Study:

To apply fluorescence lifetime correlation spectroscopy (FLCS) to resolve photophysical state dynamics in EGFP.
To quantify transition rates between distinct fluorescent states of EGFP.
To elucidate the molecular basis of these rapid transitions.

Main Methods:

Utilizing fluorescence lifetime correlation spectroscopy (FLCS).
Analyzing microsecond transition rates between fluorescent states.
Correlating spectral properties with molecular dynamics.

Main Results:

Successfully resolved the photophysical state dynamics of EGFP using FLCS.
Quantified microsecond transition rates between two fluorescent states with overlapping emission spectra.
Linked these transitions to room-temperature, angstrom-scale rotational isomerism of a nearby amino acid.

Conclusions:

FLCS is a powerful technique for studying rapid transition dynamics in complex light-emitting systems.
The study provides insights into the photophysics of EGFP at the molecular level.
This approach is applicable to a broad range of systems with multistate photophysics.