The aim of the present study in beef cattle was to investigate potential differences in follicle size and follicle wall-blood flow between cows and heifers and to compare follicle wall-blood flow between smaller and larger follicles. Cows and heifers were treated with a synchronization protocol and follicles and CLs were measured and evaluated for blood flow. Pregnancy diagnosis was performed on day 50 of the protocol. Cows had larger (P < 0.008) follicles than heifers. Cows, heifers, and pregnant and non-pregnant cows did not differ (P > 0.05) in CL diameter, CL blood flow, and plasma progesterone concentrations. Moderate correlations between follicle diameter and follicle blood flow were observed for cows (r = 0.51; P < 0.002) and heifers (r = 0.61; P < 0.0001). Pregnant cows tended (P < 0.1) to have larger follicles between 12 to 60 h before ovulation, and had larger (P < 0.05) follicles than non-pregnant cows at hour 24 before ovulation and at hour 12 before maximum values. Pregnant cows had greater (P < 0.05) follicle blood flow than non-pregnant cows at hours −36 and −24 before maximum values. Follicle blood flow was greater (P < 0.002) in the large follicles compared with the small follicles, and tended (P < 0.06) to be greater than in medium follicles. Moderate to strong correlations were found between follicle blood flow and diameter of small (r = 0.59; P < 0.002), medium (r = 0.50; P < 0.02), and large (r = 0.71; P < 0.0001) follicles. Pregnancy rates were similar (P > 0.05) among all follicle diameter categories. In conclusion, synchronized beef cows and pregnant cows had larger follicles and greater blood flow than heifers and non-pregnant cows, and follicle wall blood flow was closely associated with increasing follicle diameter.