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The purpose of this study was to examine the potential of diffusion-weighted (DW) three-dimensional (3D) MP-RAGE sequence for cerebral ischemia of a rat brain in vivo. The prototype of a DW-3D-MP-RAGE (3D-DWI) sequence was implemented at 2.0T animal experiment MR imager. The advantage of 3D imaging are that you obtain thinner and more slices with better profiles, and better signal-to-noise ratio for an equivalent slice thickness. Since 3D imaging is preferable for obtaining contiguous thin-slice MR images, image data with 3D imaging are expected to volume estimate of imaging area. Three-dimensional calculated absolute apparent diffusion coefficient (ADC) maps were compared with coronal stained (triphenyltetrazolium chloride: TTC) brain sections. From rat experimental results, the region of cerebral ischemia with 3D-DWI and the region of stained brain sections were well in agreement. This technique enables three-dimensional image display of a cerebral ischemia area, and makes it possible to measure the volume of a precise cerebral ischemia area in vivo.