Objective To explore the clinical application value of transvaginal ultrasound combined with the micro-vascular flow (MV-Flow) imaging in analyzing the ultrasound characteristics of endometrium during ovulation in women of childbearing age. Methods A total of 74 women of childbearing age who underwent transvaginal gynecological ultrasound examination in the First Medical Center of Chinese PLA General Hospital from May 2022 to July 2022 were selected. Among them, aged 22-48 (34.4±5.0) years, including 35 women ≥35 years old in the elderly group and 39 women <35 years old in the younger group. All patients with clinically diagnosed of infertility were divided into infertility group (27 cases) and healthy control group (47 cases). On the day of ovulation, the endometrial thickness, peristalsis score, volume of endometrium, pulsatility index (PI) and resistance index (RI) of uterine spiral artery, PI and RI of bilateral uterine arteries, three-dimensional energy doppler flow parameters [blood flow index (FI), vascularization index (VI), vascular blood flow index (VFI)] and endometrial microvascular bleeding vascularization index (VI_MV) were measured by the transvaginal ultrasound. The above ultrasonic indexes were compared and analyzed between the two groups. Results There was no significant difference in the history of infertility between the younger group and the older group (P>0.05). The RI of uterine spiral artery in the younger group was lower than that in the older group, while the three-dimensional energy doppler flow parameters (VI, FI, VFI) and the VI_MV of endometrium in the younger group were significantly higher than those in the older group. There was no significant difference in age stratification between infertility group and healthy control group (P>0.05). The endometrial thickness, endometrial peristalsis score, endometrial volume, endometrial three-dimensional energy doppler flow parameters (VI, FI, VFI) and the VI_MV in inferbility group were significantly lower than those in healthy control group (P<0.05). Conclusion Transvaginal ultrasound indicators can evaluate the endometrium characteristics of women of childbearing age during ovulation to some extent, especially, VI_MV can quantitatively analyze the microblood perfusion of endometrium.