OBJECTIVE: Peripheral nerve injury in the extremities usually has poor prognosis and a high disability rate even after repair because the anatomy of the peripheral nerves remains unknown. Thus, three-dimensional (3D) reconstruction of the inside of the peripheral nerve bundles is considered as a potential solution. The aim of this study was to obtain the 3D reconstruction of the tibial nerve from calcium chloride-enhanced micro-computed tomography (micro-CT) images. STUDY DESIGN: In this study the tibial nerve samples were dyed with a saturated solution of calcium chloride and scanned by micro-CT to acquire two-dimensional (2D) images. The seed images were annotated artificially, and the 2D edge contours of the nerve bundles were extracted automatically from a series of images. On this basis, the 3D model of the tibial nerve was constructed. RESULTS: The 3D model of the tibial nerve was successfully constructed. Calcium chloride enhanced the visualization in all micro-CT, and visualization was substantially improved by the image processing algorithm. The 3D renderings provided detailed visualization of the tibial nerve. Furthermore, compared to the physical slice preparation method, similarity, repeatability, and precision were improved considerably using the micro-CT scanning method. CONCLUSION: 3D texture features model reconstruction based on calcium chloride-enhanced microCT is a major breakthrough in the field of 3D reconstruction of the bundles inside the peripheral nerves and lays the basis for further research into peripheral nerve bundles. © Science Printers and Publishers Inc.