Polytetrafluoroethylene (PTFE) micro-porous membranes were prepared from PTFE fine powder through extruding, rolling, and uniaxial longitudinally stretching. In contrast to conventional planar transverse stretching, a novel 3D mold design of non-planar transverse stretching process was employed in this study to produce micro-porous structure. The morphology, membrane thickness, mean pore size, and porosity of the PTFE membrane were investigated. The results show that the non-planar transverse stretched membranes exhibit more uniform average pore diameter with thinner membrane thickness. Morphological changes induced by planar and non-planar transverse stretching for pore characteristics were investigated. The stretching conditions, stretching temperature and rate, affect the stretched membrane. Increasing temperature facilitated the uniformity of pore size and uniformity of membrane thickness. Moreover, increase in stretching rate resulted in finer pore size and thinner membrane.