Introduction: High-frequency ultrasonography enables visualization the layered structure of the skin and shows the epidermis, dermis, subcutaneous tissue and skin appendages: hair follicles, sebaceous glands, sweat glands and blood vessels. The aim of this study was to apply high-frequency ultrasonography to evaluate the structure of the vulva, vagina and cervix, and to describe their anatomy in ultrasound. Material and method: The examinations were conducted with the use of high-frequency probes: DermaView 48 MHz and Episcan 50 MHz, by three operators experienced in performing classical ultrasound scans (with 30-, 10- and 9-year experience). The study involved 50 women aged 20–80 years who reported for a periodical gynecological check-up and presented no symptoms within the reproductive system. Results: In this study, the individual layers of the skin in the area of the mons pubis, labia majora (together with hair follicles and sweat glands) and labia minora were successfully visualized in all the patients. The subepidermal low-echogenicity band was seen in the area of the mons pubic and labia majora in 40 cases. This concerned 100% of women who underwent cosmetic skin treatments. In all the patients, HFUS demonstrated the layered structure of the anterior and posterior vaginal walls, the walls of the urinary bladder and rectum, layers of the vaginal portion of the cervix and the external opening of the cervix. Conclusions: High-frequency ultrasonography offers new quality of vulvar, vaginal and cervical imaging. It can be used for evaluation of the vulva and vagina, and confirms their layered structure. It also enables accurate measurements of the thickness of the vulvar epidermis and dermis as well as the thickness of the vaginal walls.