PROBLEM TO BE SOLVED: To provide a glass substrate for formation of a lipid flat membrane, which allows anyone to reliably form a lipid flat membrane having high resistance to a high-potential load, a small in area, and stability for a long time.SOLUTION: In a glass substrate 1 for formation of a lipid flat membrane, a resist layer 2 which covers the surface and is resistant to blast is provided on one principal surface 1a of the glass substrate 1, and in the resist layer 2, an opening 2h is formed at a position of a through hole 1h to be formed in the material of the glass substrate. The glass substrate 1 is subjected to a shot blast comprising blowing a projection material toward the resist layer from above. The through hole 1h for formation of a lipid flat membrane is formed at the position of the opening 2h of the resist layer 2 of the glass substrate 1, and a high-voltage pulse discharge treatment is preferably applied to the surrounding of the opening of the through hole 1h after the shot blast step so
