• High-performance tracking of BEVs in living systems has been realized with an AIEgen. • The probe can illuminate BEVs and shows remarkable merits over traditional methods. • The spatial and temporal distributions of BEVs within host can be clearly visualized. The nano-sized, membrane-enclosed bacterial extracellular vesicles (BEVs) released by bacteria are now regarded as important mediators involved in various physiological and pathological processes of human. To elucidate their roles in bacteria-to-host communication, methods for visualizing and tracking of these nano-vesicles are crucial, but ideal means are lacking currently. In this work, we report the development of a bioprobe, DPASP, for BEV bioimaging with superior performance. DPASP can target BEVs with boosted fluorescence due to its aggregation-induced emission (AIE) attribute, which allows BEV labeling with a high signal-to-background ratio. For BEV imaging, DPASP outperforms the commercial dyes in terms of labeling efficiency, retention time, photostability, and biocompatibility. The applications of DPASP in studying the spatial and temporal distributions of E. coli -derived BEVs within living cells and mice were also demonstrated. The approach developed here provides a valuable tool to better understand the roles of BEVs in health and diseases.