Aggregation-induced emission (AIE) dye-based cross-linked fluorescent polymeric nanoparticles (FPNs) are facilely prepared via a two-step polymerization process including emulsion polymerization and subsequent anhydride cross-linking. Then, a variety of characterization methods are carried out to determine the performance of the FPNs, which show high dispersibility and strong fluorescence in an aqueous solution due to the hydrophilic carboxyl groups on the surfaces and the AIE components as the cores. Biocompatibility evaluation and cell imaging results suggest that these FPNs are biocompatible for cell imaging. More importantly, this cross-linking strategy is proven to overcome the issue of critical micelle concentration and opens the opportunity to develop more robust fluorescent bioprobes.