OBJECTIVES/GOALS: Recent scientific discoveries show that human sex determination and differentiation is a spectrum of developmental processes and that sex/gender traits and identities not always fit binarity. This study aims to determine whether the visual arts can effectively transmit these complex scientific ideas on sex and gender variance. METHODS/STUDY POPULATION: A one-hour lecture for undergraduate students enrolled in a behavioral neuroendocrinology course included 17th century Spanish paintings and representative work from contemporary LGBTQIA+ artists in photography. Pre and post self-evaluation was conducted through five multiple choice questions. Chi Square test was employed for statistical analysis, which required elimination of no responses from item analysis. Statistical significance was defined as p value < 0.05. Three artistic images and two scientific images were included in the questions. Acquisition of content-specific knowledge on diverse body configurations, differences of sex development (DSD), transsexuality, histopathology of the gonad and neuroanatomy of the hypothalamus was evaluated. RESULTS/ANTICIPATED RESULTS: Twenty students completed the online pre- and post-test. We found that a 17th century Spanish painting was effective at making the distinction between transsexuality and intersexuality (p < 0.0005). Similarly, an artistic image that reinterprets the Vitruvius Man was effective for presenting the argument that diverse body configurations does not imply clinical pathology (p < 0.0001). Last, a scientific image showing the histopathology of an ovotestis was effective for introducing undergraduate students to diagnostic criteria for DSD (p < 0.02). No significant differences were found in the use of an artistic photograph depicting a young female to male transsexual individual nor in the use of a scientific image showing the neuroanatomical localization of the hypothalamus. DISCUSSION/SIGNIFICANCE: The learning of complex scientific concepts on human sex determination and differentiation can be affected by preconceived ideas, values, and attitudes towards sex and gender variance. The visual arts can provide a familiar ground of understanding between teachers and learners to transform such preconceptions.