Bifunctional nanoprobes with both magnetic and optical contrast have been developed for ultra-sensitive brain tumor imaging at the cellular level. The nanoprobes were synthesized by simultaneously incorporating a magnetite nanoparticle cluster and fluorescence dyes into silica encapsulation by a sol-gel approach under ultrasonic treatment. The nanoprobes maintain superparamagnetic behavior at room temperature and possess enhanced transverse relaxivity and good photostability. As a glioma targeting ligand, chlorotoxin was covalently bonded to the surface of the nanoprobes. In vitro cellular uptake assays demonstrated that the nanoprobes were highly specific, taken up by human U251-MG glioma cells via receptor-mediated endocytosis. The labeled glioma cells were readily detectable by both MR imager and confocal laser scanning microscopy.