The white matter (WM) of the brain of type 2 diabetes mellitus (T2DM) patients is susceptible to neurodegenerative processes, but the specific types and positions of microstructural lesions along the fiber tracts remain unclear.In this study 61 T2DM patients and 61 healthy controls were recruited and underwent diffusion spectrum imaging (DSI). The results were reconstructed with diffusion tensor imaging (DTI) and neurite orientation dispersion and density imaging (NODDI). WM microstructural abnormalities were identified using tract-based spatial statistics (TBSS). Pointwise WM tract differences were detected through automatic fiber quantification (AFQ). The relationships between WM tract abnormalities and clinical characteristics were explored with partial correlation analysis.TBSS revealed widespread WM lesions in T2DM patients with decreased fractional anisotropy and axial diffusivity and an increased orientation dispersion index (ODI). The AFQ results showed microstructural abnormalities in T2DM patients in specific portions of the right superior longitudinal fasciculus (SLF), right arcuate fasciculus (ARC), left anterior thalamic radiation (ATR), and forceps major (FMA). In the right ARC of T2DM patients, an aberrant ODI was positively correlated with fasting insulin and insulin resistance, and an abnormal intracellular volume fraction was negatively correlated with fasting blood glucose. Additionally, negative associations were found between blood pressure and microstructural abnormalities in the right ARC, left ATR, and FMA in T2DM patients.Using AFQ, together with DTI and NODDI, various kinds of microstructural alterations in the right SLF, right ARC, left ATR, and FMA can be accurately identified and may be associated with insulin and glucose status and blood pressure in T2DM patients.© 2022. The Author(s).