Fluorescent probes for sensitive and selective detection of pH changes in live cells in visible and near-infrared channels

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We report five fluorescent probes based on coumarin-hybridized fluorescent dyes with spirolactam ring structures (A–E) to detect pH changes in live cells by monitoring visible and near-infrared fluorescence changes. Under physiological or basic conditions, the fluorescent probes A, B, C, D, and E preserve their spirolactam ring-closed forms and only display fluorescent peaks in the visible region corresponding to coumarin moieties at 497, 483, 498, 497 and 482 nm, respectively. However, at acidic pH, the rings of the spirolactam forms of the fluorescent probes A, B, C, D, and E open up, generating new near-infrared fluorescence peaks at 711, 696, 707, 715, and 697 nm, respectively, through significantly extended π-conjugation to coumarin moieties of the fluorophores. The fluorescent probes B and E can be applied to visualize pH changes by monitoring visible as well as near-infrared fluorescence changes. This helps avoid fluorescence imaging blind spots at neutral or basic pH, which typical pH fluorescent probes encounter. The probes exhibit high sensitivity to pH changes, excellent photostability, low auto-fluorescence background and good cell membrane permeability.

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©The Royal Society of Chemistry 2017. Publisher's version of record: https://dx.doi.org/10.1039/C7TB02583A

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Journal of Materials Chemistry B