摘要： Developing of two-photon materials for live-cell imaging and in vivo analysis in-deep have been received great attention, and it is still urgent so that such microscopy technique could be promoted and advanced into new progress using the powerful probs. Herein, a new arylamino fumaronitrile derivative NPAPF was synthesized and transferred as aggregation-induced emission luminogen (AIEgen) fluorescent nanoparticles (AF-NPs) via assembly technique. This AF-NPs exhibited two-photon absorption cross-section at 2.6×106 GM with 19.5% of fluorescence quantum yield. Moreover, utilizing the great potential of AF-NPs, two-photon imaging of live cells with good cytocompatibility is realized upon two-photon microscopy. By in vivo long-term tracing studies of mesenchymal stem cells, we demonstrated the tremendous advantage of AF-NPs tracer in monitoring the stem cells transplant. Therefore, our unique AF-NPs provided an efficient two-photon-absorbing probe for investigating biological mechanism and behavior, and also opened a new avenue for spatiotemporal visualization of transplanted stem cells.