In order to study potent microenvironments of malignant gliomas with a high- resolution x-ray imaging technique, an injection orthotopic glioma model was made using the Sprague-Dawley rat. Total brain tissue, taken out as an ex vivo model, was examined with diffraction-enhanced imaging (DEI) computed tomography (CT) acquired with a 35 keV monochromatic x-ray. In the convolution-reconstructed 2D/3D images with a spatial resolution of 12.5 × 12.5 × 25 µm, distinction among necrosis, typical ring-shaped viable tumors, edemas and healthy tissues was clearly observed near the frontal lobe in front of the rat's caudate nucleus. Multiple microvascular proliferations (MVPs) were observed surrounding peritumoral edemas as a tumor infiltration structure. Typical dimensions of tubular MVPs were 130 (diameter) ×250 (length) µm with a partial sprout structure revealed in the 3D reconstructed image. Hyperplasia of cells around vessel walls was revealed with tumor cell infiltration along the perivascular space in microscopic observations of mild MVP during histological analysis. In conclusion, DEI-CT is capable of imaging potent tumor-infiltrating MVP structures surrounding high-grade gliomas.