Cryopreserved mammalian sperm generally exhibit lower fertility than fresh sperm. An increasing number of antioxidants have been tested in attempts to improve semen quality, but improvements have not often been consistent or repeatable. The objective of this study was to determine if adding several different antioxidant combinations to ram sperm before cryopreservation could improve sperm binding to perivitelline membrane (PM) after thawing. Thirty ejaculates from 3 rams were split and diluted to 200 × 10⁶ sperm/ml in an egg yolk tris diluent containing different antioxidants: control, with no antioxidant; 100 μM Melatonin plus 0,05% ascorbic acid (MEL+AA); 100 μM MEL plus 90μL Trolox C (MEL+TRO); 90μL TRO plus 0,05% AA (TRO+AA); and 100 μM MEL plus 0,05% AA plus 90 μL TRO (MEL+AA+TRO). The samples were then cooled to 5°C, over 2h, and upon reaching 5°C were packaged into 0.5mL straws and frozen in static liquid nitrogen vapor for 15min before being plunged into liquid nitrogen. Straws were thawed for 30s in 37°C water and the motility and zona binding ability were determined using a CASA and fluorescence microscopy, respectively. Prior to binding analysis, sperm were stained with 35 μg/mL Hoechst 33342 for 15 min at 37°C, and then centrifuged at 400g for 5min and the sperm suspended in 1mL TALP and 5-μL aliquots (10,000 sperm prepared) were added to each PM. The membranes and sperm incubated together at 37°C for 2 h in an atmosphere of 5% of CO₂ in air, after which the PM were washed 5 times, placed onto a glass slide and examined using fluorescence microscopy at 400×. The number of sperm bound to the membrane in 6 predetermined fields of each membrane piece was counted. Data were analyzed by ANOVA using Tukey test. All antioxidants combinations maintained higher percentages of total and progressive motile cells after thawing, than control sperm (P < 0.05), with MEL+AA+TRO maintaining the highest percentage of motile sperm (49%). Sperm treated with MEL+AA+TRO exhibited the highest the number of sperm binding to the PM after thawing (177.63 cells; P < 0.05) compared with other treatments. Adding antioxidants to ram sperm before freezing increased the number of sperm surviving cryopreservation. Supported by FACEPE, CAPES and UNIVASF (Brazil).