By using molecular imprinting techniques, the Cu2+ imprinted crosslinked chitosan microspheres were prepared by using chitosan as functional monomer, Cu2+ as template ions, solidified by dilute ammonia, epoxy chloropropane crosslinked, and eluted Cu2+ by hydrochloric acid. The products were characterized by fourier transform infrared spectroscopy、X-ray diffractometer and field emission scanning electron microscopy. The physical properties such as skeleton density, water content, crosslinking degree etc of Cu2+ imprinted crosslinked chitosan microspheres were measured. The results showed that crosslinking modification can make the microspheres have porous structure and good structural stability, it also solve the acid solubility of chitosan very well, and the adsorption properties of microspheres on Cu2+ were improved. Through the orthogonal test and used table L9(34), the optimum preparation conditions of Cu2+ imprinted crosslinked chitosan microspheres were as follows: the quality of chitosan was 1.5 g, the dosage of epoxy chloropropane was 2.5 mL, crosslinked temperature was 80℃, crosslinked time was 3 h, the best adsorption property of microspheres on Cu2+ was obtained. The adsorption properties of microspheres to Cu2+ were also studied in single component system. The results showed that when the mount of microspheres was 50 mg, the initial concentration of Cu2+ was 338.65 mg/L, the pH value was 5, the adsorption capacity of Cu2+ was 72.8 mg/g.