Polydimethylsiloxane (PDMS) and glass are among the most widely used materials in biomedical and microfluidic applications. In this paper, oxygen plasma exposure was used to improve the adhesion properties of PDMS and glass. The effect of bonding quality parameters such as RF power, time of activation and oxygen flow was investigated. Bonding area and strength, two main indicators of bonding quality, were detected using manual peel and mechanical shear tests, respectively, to optimize the bonding parameters. It was observed that increase in activation time and RF power increased the bonding strength considerably. The oxygen flow had a slight influence in increasing the bonding strength. The application of this bond has also been demonstrated in PDMS-glass micropump, so this technique can be potentially applied for fabrication of PDMS-glass-based microfluidic and biomedical devices.