Abstract Background The aim of this study was to examine the implementation and sustainability of checklist use among procedural-based specialties in a pediatric hospital and to survey perceptions of checklist efficacy among staff members and physicians. Methods A modified World Health Organization procedural checklist was implemented. Percentage daily compliance was collected for all procedures over a 12-month period. Clinical staff members participated in a survey regarding the checklist. Associations among gender, clinical role, and years of experience were evaluated. Results Compliance at 12 months was significantly higher than at inception (94% vs 88%, P < .001) with average monthly compliance of 92%. Compliance deteriorated on weekends and holidays. Surgeons had more positive perceptions of using the checklist compared with nurses and anesthesiologists, independent of experience and gender ( P = .001). Conclusions The implementation of a procedural checklist can be sustained across specialties with high compliance. Off-hours utilization remains problematic. Perceptions of checklist efficacy are disparate among roles in the operating room.