Our laboratory utilizes Supported Membrane Templates (SMrT)—a versatile in vitro platform capable of generating diverse membrane topologies from planar bilayers to highly curved nanotubes—to systematically screen cell and tissue lysates for novel membrane integrity disrupting (MID) proteins. When perfused over the SMrT platform, MID activity is visualized via real-time fluorescence microscopy and characterized by distinct morphological alterations, including the budding and tubulation of planar patches or the fission of membrane nanotubes (see movies). Following the visual capture of these remodeling events, we isolate the causative molecular factors using biochemical fractionation paired with mass spectrometry, ultimately validating their conserved mechanisms and physiological functions in vivo using budding yeast and mammalian cell lines.