Objective: To investigate the optimization of preoperative preparation processes in the operating room and its application value in gastrointestinal surgery. Methods: One hundred and twenty five patients undergoing gastrointestinal surgery were selected. Before the start of surgery, 35 cases received the collaborative optimization measures that were established after discussion among anesthesiologists, surgeons and operating room nurses (optimization group), while 90 cases received the conventional approaches (conventional group). The playback data from the video surveillance cameras of the operating room of the two groups of patients were analyzed. Results: Regardless of whether dividing the operative positions or not, the preparation times from the initiation of induction of anesthesia to the start of surgery in optimization group were all significantly shorter than those in conventional group (all P<0.05). In supine position surgery, the times from initiation of induction of anesthesia to start of urinary catheterization, from the end of urinary catheterization to start of operation site disinfection, and from completion of draping to start of surgery in patients receiving optimization measures were all significantly superior to those receiving conventional approaches (all P<0.05); in lithotomy position surgery, the times from initiation of induction of anesthesia to patient positioning, from end of patient positioning to start of operation site disinfection, and from completion of draping to start of surgery in patients receiving optimization measures were all significantly superior to those receiving conventional approaches (all P<0.05). Conclusion: The established optimization measures based on multidisciplinary team collaboration can help to reduce the changeover times between processes and time for inducing anesthesia, and thereby boost the efficiency of the operating room.