Aim: This study aims to investigate the effects of 3D virtual cadaver dissection practice on learning motivation, academic achievement and self-efficacy in nursing students. Background: Anatomy is a challenging subject for nursing students due to its vast content that must be learned within a limited timeframe. Visualization-based teaching methods in anatomy education have proven to aid in understanding human structures. The use of 3D virtual cadaver dissection, facilitated by Anatomage, allows for virtual dissections of cadavers from various angles. Design: A quasi-experimental study using a non-equivalent control group pretest-posttest design. Methods: Seventy-seven first-year nursing students in an extracurricular program participated (42 experimental, 35 control). The experimental group participated in 3D virtual dissection practice, including the orientation session, once a week for 2 hours over a total of 5 weeks. To assess educational outcomes, surveys measuring learning motivation, academic achievement and self-efficacy were administered to both groups before and after the intervention. Results: For learning motivation, a statistically significant difference was observed in autonomous motivation (t = 1.68, p = .048), while no significant difference was found in controlled regulation (t = .571, p = .570). Significant differences were observed in academic achievement (t = 3.98, p < .001) and self-efficacy (t = 3.38, p = .001) between the groups. Conclusion: 3D virtual cadaver dissection practice has been demonstrated to be an effective educational method for improving autonomous learning motivation, academic achievement and self-efficacy among first-year nursing students.