To investigate the association between sleep deprivation and vestibular dysfunction by Video Head Impulse Test (vHIT). This prospective clinical trial explores the impact of acute sleep deprivation on the vestibular-ocular reflex (VOR) in medical residents. The study involved healthy physicians from diverse medical disciplines. Participants underwent vHIT assessments before and after a 26-h shift. The examinations focused solely on the right lateral semicircular canal. Participants further completed a demographics and fatigue questionnaire, including the Fatigue Severity Scale (FSS) questionnaire and a Visual Analog Fatigue Score (VAFS). The study involved 30 medical residents. Participants experienced a statistically significant decrease in VOR gain in the right horizontal semicircular canal during a 26-h shift (p < 0.01). While the FSS and VAFS questionnaires showed no significant difference before and after the shift, the analysis of ∆VOR gain indicated a statistically significant increase associated with decreased sleep time during the shift (p = 0.018, 95% Confidence Interval [0.08, 0.68]). The most substantial increase in ∆VOR occurred between 22-26 h of sleep deprivation. No significant differences were observed in ∆VOR between genders, ages, disciplines, department shifts versus emergency room shifts, or years of residency. vHIT can be used as an objective, reliable screening tool for severe sleep deprivation among physicians. The decrease in the VOR gain may indicate that vestibular function is influenced by sleep deprivation. The clinical significance of these findings is still questioned, more studies may help to assess this effect. 3 Laryngoscope, 2024.