Microfluidic devices have been used progressively in biomedical research due to the advantages they offer, such as relatively low-cost, rapid and precise processing, and an ability to support highly automated analyses. Polydimethylsiloxane (PDMS) and polymethylmethacrylate (PMMA) are both biocompatible materials widely used in microfluidics due to their desirable characteristics. It is recognized that combining these two particular materials in a single microfluidic device would enable the development of an increasingly in-demand array of new applications, including those requiring high flow rates and elevated pressures. Whereas complicated and time-consuming efforts have been reported for bonding these two materials, the robust adhesion of PDMS and PMMA has not yet been accomplished, and remains a challenge. In this study, a new, simple, efficient, and low-cost method has been developed to mediate a strong bond between PMMA and PDMS layers at room temperature in less than 5 min using biocompatible adhesive tape and oxygen plasma treatment. The PDMS-PMMA bond was hydrolytically stable, and could tolerate a high influx of fluid without any leakage. This study addresses the limitations of existing approaches to bond these materials, and will enable the development of highly sought high-pressure and high-throughput biomedical applications.