Cross-linked chitosan microspheres (40-100 microm) with smooth surface were prepared by the methods of emulsification and ethanol coagulant. FTIR results showed that the cross-linking reaction occurred on the amino groups of chitosan molecules. The swelling characteristic of chitosan microspheres was influenced by the environment pH, being generally greater at low rather than higher pH values. The coagulation properties of chitosan microspheres were evaluated by dynamic blood clotting, platelet adhesion and activation, erythrocyte adhesion, hemolysis, and protein absorption assays. Chitosan microspheres can shorten the clotting time and induce the adhesion and activation of platelets. But the shortening of clotting time by chitosan microspheres may be related to not only platelet aggregation, but also erythrocyte aggregation. Take together, chitosan microspheres may be potential use as thrombospheres.