Vianna, L. L., Pradieé, J., Santos, E. C. S., Gonçalves, A. O., Pfeifer, L. F. M., Rheingantz, M. G. T., Dode, M. A. N., Vieira, A. D., Lima, V. F. H., Correa, M. N. and Pegoraro, L. M. C. 2014. Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production. Can. J. Anim. Sci. 94: 35-42. The objective of this study was to evaluate alternatives in small volumes to conventional gradient of Percoll^® on semen quality, in vitro embryo production, sex ratio and embryo survival after vitrification. Thawed semen was randomly allocated to one of four density gradient selection methods: (1) conventional Percoll^® (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), and (4) MiniOptiprep (MO). Sperm kinetics and quality were evaluated. Use of P, MP and MI gradients did not affect sperm motility (P>0.05). However, there was a decrease in total and progressive sperm motility in MO (70.8 and 51.3% vs. 87.3 and 69.5% for P; 87.3 and 73% for MP; 92.3 and 78.8% for MI; P<0.05). The MO had lower membrane integrity compared with P, MP and MI (39.7 vs. 70.5, 72.3, 63.8%, respectively, P<0.05). The percentage of blastocysts produced was higher in MI than in MP and MO (21.1 vs. 16.1 and 16.9%, P<0.05) and similar to P (18.4%; P>0.05). Sex ratio and embryo survival after vitrification were similar among groups (P>0.05). Semen selected by Isolate and Optiprep gradient, at the concentrations and small volumes used, demonstrated similar characteristics and in vitro embryo production to conventional Percoll^® gradient.