Purpose Diffusion weighted imaging (DWI) has revealed microstructural changes in lower limb nerves in people with diabetic neuropathy. Microstructural changes in upper limb nerves using DWI in people with diabetes have not yet been explored. Methods This cross-sectional study aimed to quantify and compare the microstructure of the median and ulnar nerve in people without diabetes (n = 10), people with diabetes without distal symmetrical polyneuropathy (DSPN; n = 10), people with DSPN in the lower limbs only (DSPN FEET ONLY; n = 12), and people with DSPN in the upper and lower limbs (DSPN HANDS & FEET; n = 9). DSPN diagnosis included electrodiagnosis and corneal confocal microscopy. Tensor metrics, such as fractional anisotropy, radial diffusivity and axial diffusivity, and constrained spherical deconvolution metrics, such as dispersion and complexity, were calculated. Linear mixed-models were used to quantify DWI metrics from multiple models in median and ulnar nerves across the groups, and to evaluate potential differences in metrics at the wrist and elbow based on the principle of a distal-to-proximal disease progression. Results Tensor metrics revealed microstructural abnormalities in the median and ulnar nerve in people with DSPN HANDS & FEET, and also already in DSPN FEET ONLY. There were significant negative correlations between electrodiagnostic parameters and tensor metrics. A distal-to-proximal pattern was more pronounced in the median nerve. Non-tensor metrics showed early microstructural changes in people with diabetes without DSPN. Conclusion Compared to people without diabetes, microstructural changes in upper limb nerves can be identified in people with diabetes with and without DSPN, even before symptoms occur.