Abstract
Fluorescence-based technologies have revolutionized in vivo monitoring of biomolecules. However, significant technical hurdles in both probe chemistry and complex cellular environments have limited the accuracy of quantifying these biomolecules. Herein, we report a generalizable engineering strategy for dual-emission anti-Kasha-active fluorophores, which combine an integrated fluorescein with chromene (IFC) building block with donor-π-acceptor structural modification. These fluorophores exhibit an invariant near-infrared Kasha emission from the S1 state, while their anti-Kasha emission from the S2 state at around 520 nm can be finely regulated via a spirolactone open/closed switch. We introduce bio-recognition moieties to IFC structures, and demonstrate ratiometric quantification of cysteine and glutathione in living cells and animals, using the ratio (S2/S1) with the S1 emission as a reliable internal reference signal. This de novo strategy of tuning anti-Kasha-active properties expands the in vivo ratiometric quantification toolbox for highly accurate analysis in both basic life science research and clinical applications.
Highlights
Fluorescence-based technologies have revolutionized in vivo monitoring of biomolecules
We report a general strategy to construct activatable anti-Kasha dual-emission fluorophores based on a discovered building block, which we coin as integrated fluorescein with chromene (IFC) chromophores, wherein a spirolactone open/ closed switch can finely regulate anti-Kasha-active behavior
Among these conjugated molecules (Supplementary Figs. 1 and 2), the DCM-IFC fluorophores immediately grabbed and focused our attention when we discovered its extremely unusual dualemission
Summary
Fluorescence-based technologies have revolutionized in vivo monitoring of biomolecules. We introduce bio-recognition moieties to IFC structures, and demonstrate ratiometric quantification of cysteine and glutathione in living cells and animals, using the ratio (S2/S1) with the S1 emission as a reliable internal reference signal This de novo strategy of tuning anti-Kasha-active properties expands the in vivo ratiometric quantification toolbox for highly accurate analysis in both basic life science research and clinical applications. One effective approach to address these challenges would be to develop a single-type of probe with dual-emission signals: one serving as the internal intensity reference and the other for representing interactions with the target analyte (Fig. 1c). The ratiometric bioimaging based on the single fluorophore allows us to accurately perform quantification of various biomolecules in living cells and a mice model This de novo strategy for activatable anti-Kasha fluorophores would greatly facilitate the advancement of quantitative fluorescence imaging
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