Intracellular pH is closely related with many biological processes, including cellular proliferation, apoptosis, endocytic processes, signal transduction, and enzymatic activity. The use of fluorescent probes has become an essential method for intracellular pH detection, but existing fluorescent probes have substantial limitations, such as requiring tedious synthetic preparation, suffering from an inappropriate response range and insufficiently long emission wavelength. In this work, a red emissive two‐photon fluorescence probe based on carbon dots (pH‐CDs) is fabricated using a facile one‐pot hydrothermal method for the monitoring of intracellular pH. pH‐CDs possess a variety of superior properties, including high selectivity, excellent photostability, and low cytotoxicity. Furthermore, they exhibit a pH‐sensitive response in the range of 1.0–9.0 and a linear range of 3.5–6.5, which is desirable for tracking the pH value in living cells. It is demonstrated that the pH‐dependent fluorescence signal is regulated via switching between aggregation and disaggregation of CDs. More importantly, pH‐CDs can be successfully applied to sense and visualize pH fluctuation in cells, tissue, and zebrafish. These findings suggest that the as‐prepared pH‐CDs probe has significant potential for practical application in living systems.

A ratiometric two-photon probe was designed to visualize HClO levels in tissues, revealing the generation of HClO in the wound-repairing process of mice for the first time.

Microelectrode‐based electrochemical (EC) and photoelectrochemical (PEC) sensors are promising candidates for in vivo analysis of biologically important chemicals. However, limited selectivity in complicated biological systems and poor adaptability to electrochemically non‐active species restrained their applications. Herein, we propose the concept of modulating the PEC output by a fluorescence resonance energy transfer (FRET) process. The emission of energy donor was dependent on the concentration of target SO2, which in turn served as the modulator of the photocurrent signal of the photoactive material. The employment of optical modulation circumvented the problem of selectivity, and the as‐fabricated PEC microelectrode showed good stability and reproducibility in vivo. It can monitor fluctuations of SO2 levels in brains of rat models of cerebral ischemia‐reperfusion and febrile seizure. More significantly, such a FRET modulated signaling strategy can be extended to diverse analytes.

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