An apparatus for single-sided bilayer formation includes a first fluid chamber including a sidewall and a second fluid chamber extending through the sidewall. A barrier wall, having at least a portion defining a hydrophobicity or hydrophilicity surface property, separates the first and second fluid chambers and includes a nanopore therein across which a planar lipid bilayer (PLB) is formed. In use, an electrolyte is added to the first and second fluid chambers and a lipid/organic solvent mixture is added to the first fluid chamber to form a lipid/organic solvent layer. The electrolyte level within the first fluid chamber is adjusted such that the lipid layer is raised above the barrier wall and a PLB is formed through single-sided spontaneous formation from the first fluid chamber across the nanopore.