In this video, we will show you how we produce functional lipid bilayers that mimic biological membranes and allow for the investigation of membrane-confined processes such as they occur in neurotransmission, endo- and exocytosis. To mimic the structure and function of a biological membrane, we make use of highly ordered pore arrays in silicon chips and generate so-called pore-spanning membranes. Such membranes enable us to control and adjust parameters like lipid composition and the membrane’s mechanical properties. Proteins participating in the membrane-confined processes can be isolated and reconstituted into these pore-spanning membranes. This reconstitution allows us to address fundamental biological questions on a molecular level.