Membranes play a critical key role in many microfluidic systems, enabling diverse applications in different research fields. Our review article nicely displays an introduction into microfluidic systems applying membrane technology for analytical and organ-on-chip systems. In most cases, polydimethylsiloxane (PDMS) is used to build microfluidic chips. This is, however, accompanied by a number of limitations. For this reason, the review focuses on the integration of membranes into PDMS-free systems.