Background Nasolabial fold (NLF) reveals the aging of the face, and various treatments for NLF have been developed currently. However, most methods introduced to evaluate NLF are subjective and qualitative. Objectives This study was aimed to establish and validate a quantitative evaluation method by three-dimensional imaging, and thus accordingly to determine the relationship between quantitative measurements and wrinkle severity rating scale (WSRS) score. Methods Seventy-five Asian female volunteers were enrolled for evaluating the severity of NLF based on both WSRS score and three-dimensional imaging. Three-dimensional imaging was used to collect the quantitative measurements of NLF, including length, width, and depth. The relationship between multiple WSRS scores and quantitative measurements was compared statistically. Besides, thirty patients with moderate or severe NLF were admitted to receive hyaluronic acid injection for NLF correction. All NLFs were evaluated quantitatively preinjection and 3 months post-injection. Results The results revealed a statistically significant positive relationship between the WSRS score and the length, width, and depth of NLF, respectively. After 3 months of hyaluronic acid injection, the mean WSRS score, the length, width, and depth of NLF were significantly reduced. Conclusions Three-dimensional imaging can be used to quantitatively assess the severity of the nasolabial folds, compensating for the deficiencies of traditional scales. This technique may help improve rejuvenation procedures for the aging nasolabial fold.