PurposeWe hypothesized that the time‐dependent diffusivity at short diffusion times, as measured by oscillating gradient spin echo (OGSE) diffusion MRI, can characterize tissue microstructures in glioma patients.Theory and MethodsFive adult patients with known diffuse glioma, including two pre‐surgical and three with new enhancing lesions after treatment for high‐grade glioma, were scanned in an ultra‐high‐performance gradient 3.0T MRI system. OGSE diffusion MRI at 30–100 Hz and pulsed gradient spin echo diffusion imaging (approximated as 0 Hz) were obtained. The ADC and trace‐diffusion‐weighted image at each acquired frequency were calculated, that is, ADC (f) and TraceDWI (f).ResultsIn pre‐surgical patients, biopsy‐confirmed solid enhancing tumor in a high‐grade glioblastoma showed higher and lower , compared to that at same OGSE frequency in a low‐grade astrocytoma. In post‐treatment patients, the enhancing lesions of two patients who were diagnosed with tumor progression contained more voxels with high and low , compared to the enhancing lesions of a patient who was diagnosed with treatment effect. Non‐enhancing T₂ signal abnormality lesions in both the pre‐surgical high‐grade glioblastoma and post‐treatment tumor progressions showed regions with high and low , consistent with infiltrative tumor. The solid tumor of the glioblastoma, the enhancing lesions of post‐treatment tumor progressions, and the suspected infiltrative tumors showed high diffusion time‐dependency from 30 to 100 Hz, consistent with high intra‐tumoral volume fraction (cellular density).ConclusionDifferent characteristics of OGSE‐based time‐dependent diffusivity can reveal heterogenous tissue microstructures that indicate cellular density in glioma patients.