AbstractPhotosynthesis is a very important process for the current biosphere which can maintain such a subtle and stable circulatory ecosystem on earth through the transformation of energy and substance. Even though been widely studied in various aspects, the physiological activities, such as intrinsic structural vibration and self‐regulation process to stress of photosynthetic proteins, are still not in‐depth resolved in real‐time. Herein, utilizing silicon nanowire biosensors with ultrasensitive temporal and spatial resolution, real‐time responses of a single photosystem I‐light harvesting complex I (PSI‐LHCI) supercomplex ofPisum sativumto various conditions, including gradient variations in temperature, illumination, and electric field, are recorded. Under different temperatures, there is a bi‐state switch process associated with the intrinsic thermal vibration behavior. When the variations of illumination and the bias voltage are applied, two additional shoulder states, probably derived from the self‐conformational adjustment, are observed. Based on real‐time monitoring of the dynamic processes of the PSI‐LHCI supercomplex under various conditions, it is successively testified to promising nanotechnology for protein profiling and biological functional integration in photosynthesis studies.