Background: Research on burnout in the medical community has extensively studied the impact of mindfulness-based interventions (MBIs), which can include meditation, outdoor retreats, in-person didactics, and/or online wellness modules. However, in addition to these MBIs lacking objective, physiological measures for wellness, there has been little to no research involving virtual reality (VR) as an MBI modality for healthcare professionals in the United States. Methods: A randomized controlled intervention trial was used to study the impact of VR-based guided-meditations in the form of brief paced-breathing exercises. Heart-rate variability (HRV), a biomarker for relaxation, was measured during each session. Thirty-two participants, consisting of primarily medical students, resident physicians, and registered nurses, were recruited to complete brief guided-meditations via a VR headset or a standalone mobile app in the emergency department (ED) on-call room of a large urban academic medical center. Results: A total of 213 guided-meditation sessions were completed over the course of 4 weeks. Self-reported ratings of anxiety improved in both VR and mobile groups post-study. However, the VR group demonstrated higher intrasession HRV progress, indicating increased state of relaxation that also correlated with the number of sessions completed. Analysis by gender revealed disparity in HRV metrics between male and female VR participants. Conclusion/Application to Practice: VR-based guided meditations prove to be a feasible and accessible MBI that does not require extensive time commitment for healthcare workers. VR may be a more effective meditation platform compared with standalone mobile meditation apps, especially when used on a routine basis.