Purpose To develop a radiology-pathology coregistration method for 1:1 automated spatial mapping between preoperative rectal MRI and ex vivo rectal whole-mount histology (WMH). Materials and Methods This retrospective study included consecutive patients with rectal adenocarcinoma who underwent total neoadjuvant therapy followed by total mesorectal excision with preoperative rectal MRI and WMH from January 2019 to January 2022. A gastrointestinal pathologist and a radiologist established three corresponding levels for each patient at rectal MRI and WMH, subsequently delineating external and internal rectal wall contours and the tumor bed at each level and defining eight point-based landmarks. An advanced deformable image coregistration model based on the linearized iterative boundary reconstruction (LIBR) approach was compared with rigid point-based registration (PBR) and state-of-the-art deformable intensity-based multiscale spectral embedding registration (MSERg). Dice similarity coefficient (DSC), modified Hausdorff distance (MHD), and target registration error (TRE) across patients were calculated to assess the coregistration accuracy of each method. Results Eighteen patients (mean age, 54 years ± 13 [SD]; nine female) were included. LIBR demonstrated higher DSC versus PBR for external and internal rectal wall contours and tumor bed (external: 0.95 ± 0.03 vs 0.86 ± 0.04, respectively, P < .001; internal: 0.71 ± 0.21 vs 0.61 ± 0.21, P < .001; tumor bed: 0.61 ± 0.17 vs 0.52 ± 0.17, P = .001) and versus MSERg for internal rectal wall contours (0.71 ± 0.21 vs 0.63 ± 0.18, respectively; P < .001). LIBR demonstrated lower MHD versus PBR for external and internal rectal wall contours and tumor bed (external: 0.56 ± 0.25 vs 1.68 ± 0.56, respectively, P < .001; internal: 1.00 ± 0.35 vs 1.62 ± 0.59, P < .001; tumor bed: 2.45 ± 0.99 vs 2.69 ± 1.05, P = .03) and versus MSERg for internal rectal wall contours (1.00 ± 0.35 vs 1.62 ± 0.59, respectively; P < .001). LIBR demonstrated lower TRE (1.54 ± 0.39) versus PBR (2.35 ± 1.19, P = .003) and MSERg (2.36 ± 1.43, P = .03). Computation time per WMH slice for LIBR was 35.1 seconds ± 12.1. Conclusion This study demonstrates feasibility of accurate MRI-WMH coregistration using the advanced LIBR method. Keywords: MR Imaging, Abdomen/GI, Rectum, Oncology Supplemental material is available for this article. © RSNA, 2024.