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Related Experiment Videos

[Two-photon fluorescence from recombinant green fluorescent protein].

A Xia1, S Jiang, J Zhu

  • 1National Laboratory of Molecular and Biomolecular Electronics, Southeastern University, 210096 Nanjing.

Guang Pu Xue Yu Guang Pu Fen Xi = Guang Pu
|August 30, 2003
PubMed
Summary

Recombinant green fluorescent protein (rGFP) exhibits strong two-photon fluorescence, with changes indicating proton transfer and conformational shifts. Optimization of rGFP excitation and detection is crucial for accurate fluorescence microscopy.

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Area of Science:

  • Photophysics
  • Biophysics
  • Molecular Biology

Context:

  • Recombinant green fluorescent protein (rGFP) is a vital tool in biological imaging.
  • Understanding its photophysical properties under various excitation conditions is essential for reliable data acquisition.

Purpose:

  • To investigate the photoconversion process of rGFP using two-photon excitation.
  • To elucidate the mechanisms underlying changes in rGFP fluorescence intensity and polarization.

Summary:

  • rGFP displays strong two-photon excitation fluorescence.
  • Two-photon-induced fluorescence polarization changes suggest proton transfer between rGFP chromophore states.
  • Illumination-induced conformational changes in rGFP chromophores can block energy transfer, decreasing fluorescence intensity.

Related Experiment Videos

  • Three-photon excitation revealed fluorescence from amino acid residues, indicating blockage in rGFP.

    • Impact:

    • Findings highlight the necessity of optimizing excitation and detection parameters for quantitative fluorescence microscopy using rGFP.
    • Provides insights into the photophysical behavior of fluorescent proteins, aiding in the development of improved imaging techniques.