This study aimed to determine the effects of virtual reality (VR)-based meditation on the sleep quality, stress, and autonomic nervous system balance of nursing students. Nursing students were categorized into experimental groups I (VR-based meditation) and II (concentration meditation) and a control group. Before the study’s initiation, we measured the participants’ general characteristics, and a sleep measurement device was used to measure sleep quality. Stress levels and heart rate variability were measured before and after study completion. On the first day, all three groups slept without any intervention. On days 2–6, only experimental groups I and II implemented the intervention before sleeping. We found that the subjective sleep quality, wakefulness after sleep onset, sleep efficiency, deep sleep quality, subjective stress, objective stress, and autonomic nervous system balance of the VR meditation group were significantly better than those of the other groups. Our results reveal that the participants who underwent VR-based meditation experienced better sleep quality, lower stress levels, and improved autonomic nervous system balance compared with those in the concentration meditation and control groups. Thus, VR-based meditation effectively enhances sleep quality, lowers stress levels, and improves autonomic nervous system balance in nursing students.