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Nitric oxide (NO) is often involved in many different physiological processes including the regulation of lysosomal functions. However, it remains a great challenge to explore the variations of NO levels in lysosomes, limiting the understanding behind its biological functions in cellular signaling pathways and various diseases. Herein, a pH-activatable fluorescent probe, Rhod-H-NO , was designed and synthesized for the determination of lysosomal NO, in which the activation response model is beneficial towards getting accurate biological information. To ensure that Rhod-H-NO can accumulate effectively and exist stably in lysosomes without interference and degradation from other active species, Rhod-H-NO was engineered into the nanopores of mesoporous silica nanoparticles (MSNs) with β-cyclodextrin (β-CD) as the gatekeeper to obtain a nanolab. The nanolab was successfully applied to detect lysosomal NO in living cells and in vivo with high time and spatial resolution. This nanolab could serve as an excellent molecular tool to exploit and elucidate the function of NO at sub-cellular levels.

Engineering a nanolab for the determination of lysosomal nitric oxide by the rational design of a pH-activatable fluorescent probe