BACKGROUND/AIM: This study aimed to investigate the feasibility of a mixed reality (MR)-based hologram for intraoperative navigation in colorectal surgery. Virtual reality (VR) and MR technologies can visualize overlapping three-dimensional (3D) hologram images and real space using the wearable HoloLens2 glasses. PATIENTS AND METHODS: This study comprised 13 patients with colorectal cancer. Twelve participants had hologram images created from computed tomography (CT) between August and September 2021. One patient who underwent lateral lymph node dissection (LLND) after this period was included. A 3D hologram of the arteries, veins, and tumor was downloaded to HoloLens2 with the Holoeyes MD system and used during surgery. Hologram visibility, surgical outcome, and the NASA Task Load Index (TLX) were examined. RESULTS: A total of 2 ileocecal resections, 6 right hemicolectomies, 1 partial colectomy, 4 LLNDs, and 1 para-aortic lymph node dissection were performed safely while viewing the holograms. The mean operative duration was 421 [290-555] min, blood loss was 5 [5-15] ml, and the postoperative hospital stay was 10 [9-14] days. Regarding the TLX, the mental demand score was 30 [20-40], the physical demand score was 60 [50-67.5], the temporal demand score was 50 [40-62.5], the performance score was 15 [2.5-35], the effort score was 45 [35-62.5], the frustration score was 60 [50-65], and the weighted workload score was 34 [30.17-45.835]. CONCLUSION: Viewing a hologram in VR/MR can improve the understanding of the anatomy, which cannot be ascertained on a conventional two-dimensional monitor. Holographic guidance is a highly novel surgical concept that can potentially reduce the mental demand on surgeons.