A three-dimensional positioning device for minimally invasive surgery includes a base, two angle adjustable assemblies, an arc, a sliding assembly, a first positioning assembly, a second positioning assembly and a third positioning assembly. The base includes two first frames and two second frames. Two ends of each second frame are connected to the first frames respectively. Each first frame has a pivotal member. The angle adjustable assemblies are pivotably connected to the pivotal member of the first frame respectively. The arc includes a hollow track member. The angle adjustable assemblies are connected to two ends of the arc. The sliding assembly is mounted on the track member of the arc according to a predetermined position of a nidus. Therefore, the three-dimensional positioning device can effectively improve precision and operation more precisely in minimally invasive surgery to increase surgery success rate.
