We report on the fabrication of a microarray of supported lipid bilayers (SLBs) with different chemical compositions and demonstrate its biosensing application. The technique utilizes the phenomenon of lipid self-spreading on a patterned surface, which offers complete positional selectivity for a supported lipid bilayer. We describe the fabrication of parallel 10-mu m-wide lines, each filled with an SLB with a unique composition, at 5 mu m intervals. Structures obtained with our new technique are finer and more highly integrated than previously reported structures that employ the vesicle fusion technique on patterned surfaces. We also detected specific binding between biotin microarray is valuable for biosensing applications. and streptavidin with high contrast, indicating that the microarray is valuable for biosensing applications.