Lipid bilayer membranes found in nature are heterogeneous mixtures of lipids and proteins. Model systems, such as supported lipid bilayers (SLBs), are often employed to simplify experimental systems while mimicking the properties of natural lipid bilayers. Here, we demonstrate a new method to form SLB arrays by first forming spherical supported lipid bilayers (SSLBs) on submicrometer-diameter SiO_2 beads. The SSLBs are then arrayed into microwells using a simple physical assembly method that requires no chemical modification of the substrate nor modification of the lipid membrane with recognition moieties. The resulting arrays have submicrometer SSLBs with 3 μm periodicity where >75% of the microwells are occupied by an individual SSLB. Because the arrays have high density, fluorescence from >1000 discrete SSLBs can be acquired with a single image capture. We show that 2-component random arrays can be formed, and we also use the arrays to determine the equilibrium dissociation constant for cholera toxin binding to ganglioside GM1. SSLB arrays are robust and are stable for at least one week in buffer.
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