Three-dimensional (3D) visualization of neuroanatomy can be challenging for medical students. This knowledge is essential in order for students to correlate cross-sectional neuroanatomy and whole brain specimens within neuroscience curricula and to interpret clinical and radiological information as clinicians or researchers. This study implemented and evaluated a new tool for teaching 3D neuroanatomy to first-year medical students at Boston University School of Medicine. Students were randomized into experimental and control classrooms. All students were taught neuroanatomy according to traditional 2D methods. Then, during laboratory review, the experimental group constructed 3D color-coded physical models of the periventricular structures, while the control group re-examined 2D brain cross-sections. At the end of the course, 2D and 3D spatial relationships of the brain and preferred learning styles were assessed in both groups. The overall quiz scores for the experimental group were significantly higher than the control group (t(85) = 2.02, P less than 0.05). However, when the questions were divided into those requiring either 2D or 3D visualization, only the scores for the 3D questions were significantly higher in the experimental group (F[subscript 1,85]= 5.48, P = 0.02). When surveyed, 84% of students recommended repeating the 3D activity for future laboratories, and this preference was equally distributed across preferred learning styles ([chi][superscript 2] = 0.14, n.s.). Our results suggest that our 3D physical modeling activity is an effective method for teaching spatial relationships of brain anatomy and will better prepare students for visualization of 3D neuroanatomy, a skill essential for higher education in neuroscience, neurology, and neurosurgery. (Contains 3 tables and 3 figures.)