Background: Acupoint CV23 is one of the most commonly used acupoints for the treatment of post-stroke dysphagia and tongue pain. However, care must be taken during the procedure to consider the position of glands and blood vessels in the subcutaneous space of the acupoint. Needling depths to the geniohyoid muscle reportedly range from 0.4 to 3.3 cm. Using ultrasound imaging, we aimed to observe the anatomical characteristics around acupoint CV23 to derive a safe needling depth.Methods: Ultrasound images of acupoint CV23 accessed from the Standard Ultrasound Image of Acupoint database were retrospectively analyzed for 30 participants aged in their 20s and 30s (15 male, 15 female), and the depth from the skin to the geniohyoid muscle was measured. Correlations between the needling depth and anthropometric factors (such as neck circumference) were analyzed.Results: The average needling depth to the geniohyoid muscle was 1.59 ± 0.49 cm (male; 1.43 ± 0.52 cm, female, 1.75 ± 0.42 cm, <i>p</i> = 0.03). The geniohyoid muscle, anterior digastric muscles, and genioglossus muscle were observed in the subcutaneous area of acupoint CV23, and a risk of sublingual gland damage needs to be considered for oblique insertion of the acupuncture needle. No statistically significant correlations between the needling depth and anthropometric factors were observed.Conclusion: Acupoint CV23 has a relatively shallow needling depth, and considering the presence of blood vessels and glands in the sublingual space, visualizing the surrounding anatomical structures using ultrasound was helpful to ensure safe needling practice.