A ratiometric fluorescent nanoprobe (CDs-Rho), synthesized through the simple covalent amide linkage between carbon dots (CDs) and pH-sensitive rhodamine dye (Rho), was designed for the precise sensing and imaging of extremely alkaline environments. The sensing mechanism involves the opposite pH-dependent fluorescence changes in CDs and Rho, respectively, coupled with pH-regulated FRET efficiency from CDs to Rho. The nanoprobe features a wide pH response window from pH 7.0 to 12.0 with a pK value of 11.3 and shows high sensitivity, robust anti-interference capability, and high reversibility. Moreover, the significant shifts in emission wavelength following the pH fluctuations result in two well-separated emission signals, thus ensuring the visualization of reversible and distinct color changes (from green to red) during in vivo fluorescence imaging. This work furnished a facile protocol that contributes to the advancement of a novel method for the accurate sensing and imaging of extreme alkaline environments.
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