The most common methods for three-dimensional reconstruction of peripheral nerve fascicles include histological and radiology techniques. Histological techniques have many drawbacks including an enormous manual workload and poor image registration. Mi-cro-magnetic resonance imaging (Micro-MRI), an emerging radiology technique, has been used to report results in the brain, liver and tumor tissues. However, micro-MRI usage for obtaining intraneural structures has not been reported. The aim of this study was to pres-ent a new imaging method for three-dimensional reconstruction of peripheral nerve fascicles by 1T micro-MRI. Freshly harvested sciatic nerve samples from an amputated limb were divided into four groups. Two different scanning conditions (Mannerist Solution/GD-DTPA contrast agent, distilled water) were selected, and both T1 and T2 phases programmed for each scanning condition. Three clinical sur-geons evaluated the quality of the images via a standardized scale. Moreover, to analyze deformation of the two-dimensional image, the nerve diameter and total area of the micro-MRI images were compared after hematoxylin-eosin staining. The results show that rapid mi-cro-MRI imaging method can be used for three-dimensional reconstruction of the fascicle structure. Nerve sample immersed in contrast agent (Mannerist Solution/GD-DTPA) and scanned in the T1 phase was the best. Moreover, the nerve sample was scanned freshly and can be recycled for other procedures. MRI images show better stability and smaller deformation compared with histological images. In conclusion, micro-MRI provides a feasible and rapid method for three-dimensional reconstruction of peripheral nerve fascicles, which can clearly show the intal structure of the peripheral nerve.