The prognosis of patients with esophageal carcinoma is heavily dependent on the histopathological staging of the carcinoma. Accurate preoperative histopathological staging is important for the treatment options ^{[1, 2]}.However, the common examination modalities are extremely difficult to identify the preoperative staging precisely due to the deep location of the esophagus, artifacts caused by the respiratory and cardiac motion. Magnetic resonance imaging (MRI) was reported to evaluate the esophageal layers invasion in vitro and demonstrated that high-resolution T2-weighted imaging can clearly depict 8 layers of the esophagus which can provide essential information of the carcinoma invasion ^[3]. However, the experimental specimens were after 10% formalin fixed within 24 hours ex vivo, which might cause the signal changes of the esophageal layers and carcinoma. Furthermore, the esophageal specimens were imaged along the long axis of the esophagus and the specimens were sectioned longitudinally which were not consistence with the normal physiological and anatomical state. The purpose of this study was to prospectively establish the MRI signal characteristics of the normal esophageal wall without formalin fixation and to assess the diagnostic accuracy of high-resolution MR scanner for depicting the depth of esophageal wall invasion by making the correspondence with certain histopathological slice. From March 2014 to August 2015, 59 patients with esophageal squamous cell carcinoma were potentially eligible for this prospective study. Finally 27 patients were recruited in this study, including 19 men (mean age, 62.58±8.90 years; range, 40-78 years) and 8 women (mean age, 63.00 ±7.35 years; range, 50-74 years) and went through the experiment following the workflow shown in Fig1.MR imaging was performed using a 3.0-T MRI system (Discovery MR750, GE Healthcare, Milwaukee, USA) with a 4 channel 3.5 inch animal coil. High-resolution T2-weighted MR images were obtained with the TR = 5500 ms, TEeff = 85.0 ms, slice thickness = 3.0 mm with no gap, field of view = 60 ×36 mm², matrix size = 512 ×352, flip angle = 111°, NEX=14, resulting in a voxel volume around 0.1×0.1×3 mm³ (0.03 mL). The total scan time was about 23 minutes. After the MR scanning, the specimens were fixed with formalin and stained with hematoxylin-eosin (H-E). The specimens were sectioned transversely with a 5-um-slices. The MR images were interpreted by two experienced radiologists independently, who were blinded to the results of the histopathological examinations. When the radiologists could not fully agree on the findings, a consensus was achieved by discussion. The MR diagnostic staging of the 27 esophageal carcinomas were compared with the histopathological examinations which were served as the standard of reference. The correlation between the MR images and histopathological slices were defined by the tumor’s contour or the adjacent lymph nodes. The Kappa test was used to evaluate the MR findings and the histopathological results agreement plus the interobserver agreement. Corresponding to the histological results, high-resolution MRI can clearly depict the normal esophagus as consisting of eight layers (Fig.2). The signal characteristics of each layer was shown in Table 1. MR images correctly diagnosed 4 of 5 mucosa invasion, 6 of 7 submucosa invasion, all 10 lesions with muscularis propria invasion and 5 lesions with adventia invasion (Table 2). Table 3 showed the performance of high-resolution T2WI images with respect to the assessment of invasion of the different esophageal layers. The diagnostic accuracy of mucosa invasion, submucosa invasion, muscularis propia invasion and adventia invasion were 96%, 93%, 100% and 100%, respectively. And the sensitivity and specificity were 95-100% and 80-100 %.A k value of 0.898 was obtained for the correlation between the high-resolution MRI and histopathological findings. Interobserver agreement between two radiologists was excellent (k value, 0.847). Comparing with the T1WI images, high-resolution T2WI images can clearly depict the normal esophagus as consisting of eight layers by distinguishing of signal intensity of each layer. Furthermore, precise stagement and excellent consistency with the histopathological findings may provide a good potential for the further study of esophageal carcinoma in vivo . High-Resolution MRI can provide clear delineation of the esophageal wall layers in vitro with good diagnostic sensitivity, specificity and accuracy for assessing esophageal carcinoma invasion.