Hyperbranched polymers, as a unique highly branched polymers with low viscosity, simple synthesis, mixability and high reactivity, have received intensive attention and applied to many fields of resin modification, catalysts, and film materials. In this paper, hyperbranched polymers (P1 and P2) are designed and developed by the thiol-yne "click" reaction with controlled molar ratio of reactant. After that, lanthanide ions are coordinated with P1 and P2 to prepare new hyperbranched polymer fluorescent products (P1-Ln 3+ and P2-Ln 3+ ). P1-Ln 3+ and P2-Ln 3+ exhibit excellent fluorescent properties, superb stability, and economic feasibility. Moreover, bioimaging experiments clearly exhibit that P1-Ln 3+ and P2-Ln 3+ has great potential in further biological applications.