Thermoplastic polyurethane (TPU) and silicon tooling with microscale features on its surface was employed to investigate the impact of three factors on the quality of injection molded microscale features: (1) optimized process parameters, (2) use of a more flexible thermoplastic material, and (3) used as an antistiction coating. The molded parts and tooling surface were characterized by atomic force, confocal, and scanning electron microscopy. Although both improved filling of the tooling trenches, higher mold temperatures significantly enhanced replication, but faster injection velocities contributed moderately to replication quality. With medium aspect ratio (2.3:1) trenches, the antistiction coating doubled depth ratios, enhanced the edge definition and flatness of the features, and significantly reduced tearing of the features during ejection. The flexibility of the TPU permitted easier part ejection and left less polymer residue on the tooling surface in comparison to polycarbonate and other thermoplastic polymers. POLYM. ENG. SCI., 50:411-419, 2010. (C) 2009 Society of Plastics Engineers