Abstract: There is provided a method of preparing organic-solvent-free artificial lipid membranes formed over microapertures of a substrate and a substrate for holding the artificial lipid membranes. The method of preparing artificial lipid membranes over microapertures of a substrate includes forming a lipid pattern 5 on the perimeter of microapertures 4 formed in a substrate 1, swelling the lipid pattern 5 by an electroformation method, fusing the swelled lipid pattern, and further swelling the fused lipid pattern for each microaperture to prepare dome-shaped organic-solvent-free lipid membranes 7.