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Updated: Apr 14, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
Published on: December 11, 2021
Dual fluorescence through Kasha's rule breaking: an unconventional photomechanism for intracellular probe design
Giuseppe Brancato1, Giovanni Signore2, Paolo Neyroz3
1†Scuola Normale Superiore, Piazza dei Cavalieri 7, I-56126, Pisa, Italy.
Abstract:
Dual fluorescence is an anomalous photophysical phenomenon observed in very few chromophores in which a two-color radiative process occurs that involves two distinct excited electronic states. To date its observation was linked either to electronic rearrangement of an excited fluorophore leading to two conformers with distinct emissive properties, or to a photochemical modification leading to different fluorescent species. In both cases, emission originates from the lowest excited state of the resulting molecular configurations, in line with the so-called Kasha's rule. We report here a combined theoretical and spectroscopic study showing, for the first time, an anti-Kasha dual-emission mechanism, in which simultaneous two-color emission takes place from the first and second excited state of a coumarin derivative. We argue that the observed environmental sensitivity of this peculiar optical response makes the present compound ideally suited for biosensing applications in living cells.
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