Abstract: Disclosed in the present disclosure is a manufacturing method for a 3D microelectrode. The manufacturing method includes the following steps: (1) manufacturing a 3D model of a 3D microelectrode; (2) pouring a flexible material into the 3D model, and performing demolding so as to form a flexible mold having a cavity, wherein the cavity of the flexible mold can be fitted to the 3D model; (3) performing silanization treatment on the flexible mold, then pouring a flexible material into the surface of the flexible mold having the cavity, and performing demolding so as to form a flexible 3D microelectrode substrate; and (4) manufacturing a conductive layer on the flexible 3D microelectrode substrate so as to form the 3D microelectrode. In the present disclosure, a 3D microelectrode having an ultrahigh microcolumn height can be manufactured by using a 3D printing technology and a two-time mold-reversing method.