To develop biocompatible composite microspheres for novel hemostatic use, we designed and prepared a novel biomaterial, composite microspheres consisting of carboxymethyl chitosan, sodium alginate, and collagen (CSCM). The ultra-structure of CSCM was investigated by scanning electron microscopy assay. In hemostatic function experiment, it was found that CSCM could facilitate platelet adherence, platelet aggregation, and platelet activation in vitro. Besides, the maximum swelling of CSCM submerged in PBS for 50 min was over 300% of that exhibited by commercial hemostatic compound microporous polysaccharide haemostatic powder (CMPHP). In addition, CSCM exhibited good biodegradability and non-cytotoxicity. These results demonstrated that CSCM may be useful in platelet plug formation, and this study would provide important information for further research on hemostasis experiment in vivo.