The purpose of this study was to evaluate ten commercially available latex, powder-free surgical gloves and four commercially available non-latex, powder-free surgical gloves using standardized, reproducible biomechanical parameters that included glove thickness, puncture resistance, and glove donning force. For all gloves tested, with one exception (Neolonβ’ PF), puncture resistance increased for double-gloves as compared to single-gloves. In addition, single-glove thickness was not a reliable determinant of puncture resistance for either latex or non-latex gloves. For the latex gloves, the Ultrafree double and single-gloves exhibited the highest puncture resistance. The glove donning forces for the Biogelβ’ M and Biogel Sensorβ’ single-gloves were the lowest. In contrast, the Biogel Revealβ’ and Encoreβ’ Ultra-Thick exhibited the lowest double-glove donning forces. On the basis of these performance tests of latex gloves, the surgeon should consider the Biogel Reveal as well as the Ultrafree gloves for their latex double-glove system. For the non-latex gloves, the Pure Advantage Nitrileβ’ glove had the highest puncture resistance for one layer and two layers of glove material. The thin Pure Advantage Nitrile glove was the most resistant to glove puncture. The Pure Advantage Nitrile glove had low glove donning forces for both single-glove donning configurations and both double-glove donning configurations. Consequently, we recommend the Pure Advantage Nitrile glove as the powder-free, non-latex, double-glove system.