Patterned supported lipid bilayers (SLBs) provide a model system for studying fluid lipid bilayers and transmembrane proteins in an array format. SLB arrays self-assemble on patterned self-assembled monolayers (SAMs) consisting of hexadecanethiol and glycol-terminated regions. While the mechanism of SLB formation on glass has been studied extensively, the formation of SLBs on other substrates is not necessarily well understood. Moreover, SLB arrays on patterned SAMs represent an intriguing system, since lipid vesicles do not adhere to glycol-terminated monolayers. Here, we utilize surface plasmon resonance imaging (SPRi) and kinetic analysis to examine the mechanism of SLB formation on the glycol-terminated regions of patterned SAMs and supported lipid monolayer (SLM) formation on alkyl-terminated regions of patterned SAMs. We determine that vesicles rupture to form a patterned SLB through a two-step mechanism that is dependent upon vesicle attachment at the interface of the two regions of the patterned monolayer.