Background: With the emergence of various new skin-lightening products, there is an urgent need to scientifically evaluate the efficacy and toxicology of these products, and provide scientific guidance for their use based on physiological differences between individuals. Visualized imaging methods and quantitative evaluation criteria play key roles in evaluating the efficacy of skin-lightening products. In order to quantify the changes in the multilayered morphology and endogenous components of human skin before and after the use of lightening products, high-resolution three-dimensional (3D) imaging of human skin is required.Methods: In this study, photoacoustic microscopy (PAM; SSPM-532, Guangdong Photoacoustic Medical Technology Co., Ltd.) was used to capture the morphological structures of human skin and reveal skin components quantitatively. The efficacy and safety of skin-lightening products were evaluated by measuring skin melanin concentration and observing skin morphology. The melanin concentration in the epidermis was obtained by examining the linear relationship between photoacoustic (PA) signals. Further, the epidermal thickness and the melanin distribution were obtained in the cross-sectional (x-z) and lateral (x-y) images.Finally, the efficacy of skin-lightening products was evaluated according to the concentration and distribution of melanin in the epidermis, and the safety of cosmetics was assessed by observing the vascular morphology in the dermis.Results: PAM noninvasively could assess the multilayered morphological structures of human skin, which allowed for quantification of epidermal thickness and melanin concentration of different skin sites. Based on this, the efficacy and safety of skin-lightening products in multilayer structures were quantitatively evaluated.Conclusions: As a quantitative imaging method, PAM, has the potential to accurately evaluate the use of skin-lightening products. The method can also be extended to assessments within the larger field of aesthetic medicine.