Time-Resolved Monitoring of Intracellular Processes with a Cyclical On-Off Photoswitchable Nanoprobe

Fluorescent microscopic imaging with the help of small-molecule probes (chemoprobes) is one of the most feasible approaches for noninvasive sensing of intracellular molecules. However, the "always on" property of current chemoprobes failed to achieve time-resolved monitoring. Here, we report the development of a supramolecular nanoassembling strategy to integrate multiple functions on one nanoscale probe (nanoprobe) with a cyclical on-off switchable sensing ability. The reversal of the nanoprobe can be rapidly achieved by converting the single-wavelength near-infrared (NIR) laser to two-way emissions by a lanthanum nanoparticle core that is sensitive to the light power density. Through regulating the NIR power density, the azobenzene derivative, which was doped in the surface of the lipid bilayer of the nanoprobe, can act as an "impeller" and "brake" for bio-benign activation and deactivation, respectively, of the nanoprobe in biological applications. A reduced nicotinamide adenine dinucleotide nanoprobe was constructed as the model to demonstrate precise and time-resolved monitoring of intracellular processes including cancerous glycolysis and ligand-induced enzymatic processes. We envision that this cyclical on-off switchable nanoprobe strategy will hold great promise for endowing universal chemoprobes with high precision and temporal resolution.