The Fe3O4@SiO2@vmSiO(2) microspheres with ordered mesochannels and large inter-lamellar void were successfully prepared through stepwise solution-phase interface deposition. Fe3O4 nanoparticles were coated with SiO2 via the Stober method, and they were further coated with mesoporous SiO2 using aggregation of cetyltrimethylammonium chloride as template to prepare Fe3O4@SiO2@vmSiO(2). The Fe3O4@SiO2@vmSiO(2) microspheres show a well-defined core-shell structure with high magnetization (similar to 30.9 emu g(-1)), ordered mesochannel (similar to 6.8 nm in diameter), and inter-lamellar void (similar to 30 nm). Laccase (LAC) was immobilized on a modified Fe3O4@SiO2@vmSiO(2) microsphere by covalent attachment and stabilized onto the glassy carbon electrode (GCE) surface (Fe3O4@SiO2@vmSiO(2)-LAC/GCE) in the fabrication of novel immobilized LAC biosensors for monitoring dopamine (DA). The electrochemical properties of the biosensor were investigated with electrochemical impedance spectroscopy and cyclic voltammetry. The immobilized LAC biosensor possesses good DA electrocatalytic activity with a linear range of 1.5-75 mu mol L-1 and low detection limit of 0.177 mu mol L-1 and shows strong anti-interference ability and excellent selective determination of DA that coexists with ascorbic acid. The immobilized LAC biosensor was also used to detect DA in pharmaceutical injection. The recoveries of 98.7-100.5% were obtained for the samples, which illustrate great potential for practical application.