Cows with two waves of follicular growth during the estrous cycle yield follicles that are older and larger at ovulation compared with cows having three waves. The objectives of the current research were 1) to compare fertility in cows with two or three follicular waves and 2) to examine associations between luteal function, follicular development, and fertility after breeding. Follicular waves were monitored by ultrasonography during the estrous cycle before insemination in 106 dairy cows. Fewer cows had three follicular waves before next estrus and ovulation than two waves (P < 0.01; 30% vs 68%, respectively), but pregnancy rate was higher (P = 0.058; 81 vs 63%, respectively). Cows with two waves had shorter estrous cycles (P < 0.01), with the ovulatory follicle being both larger (P < 0.05) and older (P < 0.01). In cows with three waves, luteal function was extended (P < 0.05) and the peak in plasma progesterone occurred later (P < 0.05) in the estrous cycle compared to two wave cows. Considering cows that became pregnant, luteal phase length was shorter (P < 0.05) during the estrous cycle preceding insemination than for nonpregnant cows. In conclusion, fertility was greater in lactating cows inseminated after ovulation of the third-wave follicle that had developed for fewer days of the estrous cycle as compared with two-wave cows.