Magnetic resonance imaging (MRI) has played little role for the study of peripheral nerve disease. However, recent technological advances in MRI have provided us more information about neural microstructure and higher resolution in peripheral nerves. The aim of this study is to evaluate whether diffusion tensor imaging (DTI) in MRI can detect peripheral nerve abnormalities in patients with type 2 diabetes (T2D). In this prospective, single-center study, 33 T2D patients (mean age, 60.5 ± 7.0 yr; 16 M/ 17F) and 12 healthy controls (61.8 ± 5.3 yr, 5M / 7 F) were included. All T2D patients underwent Michigan Neuropathy Screening Instrument questionnaire and quantitative sensory testing. MRI including DTI and axial T2-weight Dixon sequence was performed for each participant. DTI parameters of the tibial nerves such as fractional anisotropy (FA) and diffusivity (mean (MD), axial (AD), and radial (RD)) were calculated. FA of the tibial nerves was significantly lower in T2D patients than healthy controls at both level 1 (0.42 ± 0.07 vs. 0.57 ± 0.09, P<0.001) and level 2 (0.44 ± 0.07 vs. 0.55 ± 0.08, P<0.001). AD was also significantly lower in T2D patients than controls at both level 1 (1.97 ± 0.43 vs. 2.40 ± 0.63, P<0.05) and level 2 (2.18 ± 0.37 vs. 2.59 ± 0.47, P<0.05). RD was significantly higher in T2D patients than controls at level 1 (0.97 ± 0.21 vs. 0.83 ± 0.13, P<0.05). However, there were no differences in DTI parameters between T2D patients with peripheral neuropathy (PN) and those without PN. In summary, we have demonstrated that DTI can detect microstructural alterations of peripheral nerves in T2D patients with PN as well as those without PN, suggesting that the structural nerve damage can occur before the development of PN in T2D patients.