In a multi optical probe confocal imaging system utilizing a microlens arrays as an objective lens, a high numerical aperture is required to improve resolving power. Glass microlens arrays are suitable for high-resolution imaging since they provide outstanding optical properties with a high refractive index. We demonstrated the rapid fabrication of microlens arrays on a high refractive index optical glass substrate via laser assisted thermal imprinting. The optical performance of the fabricated glass microlens arrays were evaluated and compared to that of a polymer microlens. In contrast to the polymer, the real image afforded by, and the calculated resolution of, the imprinted glass microlens arrays were significantly better, at about 0.73 µm compared to the polymer (∼1.56 µm). Our results reveal the considerable potential of direct thermal imprinting as a rapid, single-step, low cost fabrication method for replication of glass microlens array of high dimensional accuracy affording excellent optical performance.