Background: Because of the severe deficiency of global anesthesia workforce, anesthesiologists are forced to face work overload and more night shifts, which can disturbe the biological rhythm and cause major stress and depression, causing negative even devastating outcomes for both themselves and the patients eventually. Virtual reality (VR) as a new measure to reduce stress and pain for patients, has been widely used in biomedical fields.The purpose of the study is to evaluate the potential effectiveness of VR technology in reducing the stress of night-shift anesthesiologists.Methods: In this randomized controlled, cross-over design, single-center clinical trial, a total of 30 anesthesiologists are enrolled and randomized in a 1:1 allocation to either the VR immersion group (intervention group) or the routine night-shift group (control group) with a washout of 1 week. Anesthesiologists in the intervention group undergo VR immersion for twice while anesthesiologists in the control group will not watch VR videos during the night shift. The primary outcome is the difference between two groups about the score of NASA-TLX scale. Secondary outcomes include the score of CPSS, VAS and MFI-20 scales, satisfaction degree of the participants, heart rate (HR), blood pressure (BP), the incidence of arrhythmia as well as incidence of chest tightness, headache, palpitations.Discussion: It is unknown whether the use of VR technology during the night shift can reduce the stress of night-shift anesthesiologists. With the widely use of VR technology, a positive result of the trial could spur the hospital to apply the technology to reduce the stress of night shift doctors in every department as well as offer a relatively relaxed working environment.Trial registration: ChiCTR2000031025 on March 21, 2020.