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Uncovering the Hidden Excited State toward Fluorescence of an Intracellular pH Indicator Public Deposited

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https://ir.library.oregonstate.edu/concern/articles/1r66j6353

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  • Intracellular pH (pH(i)) imaging is of paramount importance for life sciences. In this work, we implement the ultrafast electronic and stimulated Raman spectroscopies to unravel the fluorescence mechanism of an excitation-ratiometric pH(i) indicator in basic aqueous solution. After photoexcitation of the pH(i) indicator HPTS, a hidden charge-transfer (CT) state following the locally excited (LE) state is uncovered as an essential step prior to fluorescence and this LE -> CT transition is gated by ultrafast solvation dynamics. A 835 cm(-1) intermolecular vibrational mode is identified to potentially facilitate the CT-state formation on the 700 fs time scale. Dynamic correlation with the other excited-state Raman marker bands suggests that the transition between transient electronic states is aided by solvation events mostly in the molecular plane of HPTS. These vivid structural dynamics insights can enable the rational design of more efficient and bright pH(i) indicators in an H-bonding environment with controllable properties.
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  • 9
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  • 1948-7185

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