Moisturizing creams and lotions are commonly used in daily life for beauty and treatment of different skin conditions such as dryness and wrinkling, and ultrasound stimulation has been used to enhance the delivery of ingredients into skin. However, there is a lack of convenient methods to study the effect of ultrasound stimulation on lotion absorption by skin in vivo. Ultrasound biomicroscopy was adopted as a viable tool in this study to investigate the effectiveness of ultrasound stimulation on the enhancement of lotion delivery into skin. The forearm skin of 10 male and 10 female young subjects was tested at three different sites, including two lotion treatment sites with (Ultrasound Equipment - UE ON) and without (UE OFF) ultrasound stimulation and a control site without any lotion treatment. 1MHz ultrasound with a duty cycle of 1.7%, a spatial peak temporal peak pressure of 195kPa and an average power of 0.43W was used for the stimulation. The skin thickness before, immediately after (0min), and 15 and 30min after the treatment was measured by an ultrasound biomicroscopic system (55MHz). It was found that the skin thickness significantly increased immediately after the lotion treatment for both UE ON (from 1.379±0.187mm to 1.466±0.182mm, p<0.001) and UE OFF (from 1.396±0.193mm to 1.430±0.194mm, p<0.001) groups. Further comparison between the two groups revealed that the skin thickness increase of UE ON group was significantly larger than that of UE OFF group (6.5±2.4% vs. 2.5±1.3%, p<0.001). Furthermore, it was disclosed that the enhancement of lotion delivery by ultrasound stimulation was more effective for the female subjects than the male subjects (7.6±2.3% vs. 5.4±2.0% immediately after treatment, p=0.017). In conclusion, this study demonstrated that ultrasound biomicroscopy was a feasible method for studying the effectiveness of lotion treatment in vivo, and ultrasound stimulation was effective to enhance the rate of lotion absorption into skin.