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Multiphoton switching dynamics of single green fluorescent proteins
G Chirico1, F Cannone, A Diaspro
1INFM and Department of Physics, University of Milano Bicocca, Piazza della Scienza 3, 20126 Milano, Italy.
Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics
|November 5, 2004
Summary
Researchers demonstrated multi-photon photo-switching in a green fluorescent protein (GFP) mutant, revealing two distinct fluorescent states. This discovery offers new possibilities for advanced microscopy and optical data storage.
Area of Science:
- Biophysics
- Photochemistry
- Microscopy
Background:
- Green fluorescent proteins (GFPs) are widely used in biological imaging.
- Photochromic GFPs offer dynamic control over fluorescence.
- Understanding photo-switching mechanisms is crucial for advanced applications.
Purpose of the Study:
- To demonstrate multi-photon driven photo-switching in a GFP mutant.
- To investigate the distinct fluorescent states and their optical properties.
- To explore potential applications in microscopy and data storage.
Main Methods:
- Single-protein investigation techniques.
- Multi-photon excitation spectroscopy.
- Characterization of two-photon cross-section bands.
Main Results:
- Photo-switching between dark and bright states of a GFP mutant was achieved.
- Two distinct bright states with specific two-photon cross-section bands (780 nm and 870 nm) were identified.
- Independent on/off switching of fluorescence for these two species was demonstrated.
Conclusions:
- A novel photoconversion pathway for photochromic GFPs was uncovered.
- The findings suggest significant applications for multi-photon confocal microscopy.
- Potential utility in optical data-storage architectures was highlighted.