This work describes a novel microfluidic method to prepare monodispersed chitosan microspheres by using the solvent extraction method. Our strategy is that a chitosan/acetic acid aqueous solution is emulsified in an organic phase containing the extractant by using the co-flowing shear method in a co-axial microfluidic device. The formed droplets are in situ solidified within a synthesizing channel by the extraction of acetic acid from the chitosan aqueous droplets to the organic solution. Based on this approach, the size of chitosan microspheres can be successfully controlled from 100 mum to 700 mum in diameter with a variation of less than 4%. Furthermore, high loading efficiency (>95%) of Bovine serum albumin (BSA) can be in situ encapsulated. The present method has the advantages of actively controlling the droplet diameter, narrow size distribution, good sphericity, and having a simple and low cost process, with a high throughput. This approach for the preparation of chitosan microspheres will provide many potential applications for pharmaceutical area.