INTRODUCTION: To describe changes in workplace physical activity, and health-, and work-related outcomes, in workers who transitioned from a conventional to an 'activity-permissive' workplace. METHODS: A natural pre-post experiment conducted in Vancouver, Canada in 2011. A convenience sample of office-based workers (n=24, 75% women, mean [SD] age = 34.5 [8.1] years) were examined four months following relocation from a conventional workplace (pre) to a newly-constructed, purpose-built, movement-oriented physical environment (post). Workplace activity- (activPAL3-derived stepping, standing, and sitting time), health- (body composition and fasting cardio-metabolic blood profile), and work- (performance; job satisfaction) related outcomes were measured pre- and post-move and compared using paired t-tests. RESULTS: Pre-move, on average (mean [SD]) the majority of the day was spent sitting (364 [43.0] mins/8-hr workday), followed by standing (78.2 [32.1] mins/8-hr workday) and stepping (37.7 [15.6] mins/8-hr workday). The transition to the 'activity-permissive' workplace resulted in a significant increase in standing time (+18.5, 95% CI: 1.8, 35.2 mins/8-hr workday), likely driven by reduced sitting time (-19.7, 95% CI: -42.1, 2.8 mins/8-hr workday) rather than increased stepping time (+1.2, 95% CI: -6.2, 8.5 mins/8-hr workday). There were no statistically significant differences observed in health- or work-related outcomes. DISCUSSION: This novel, opportunistic study demonstrated that the broader workplace physical environment can beneficially impact on standing time in office workers. The long-term health and work-related benefits, and the influence of individual, organizational, and social factors on this change, requires further evaluation.