We sought to evaluate a three-dimensional (3D) whole-brain perfusion technique based on echo-shifting (PRESTO) for its performance in evaluation of acute stroke. Twenty-six patients were scanned within 6 hours after onset of hemispheric symptoms, and the results were compared with results of diffusion-weighted imaging (DWI) and digital subtraction angiography (DSA). The signal-to-noise ratio of the images was 61 +/- 3 pre-contrast and 47 +/- 3 at the bolus peak. Brain coverage on perfusion parameter maps was 95% +/- 2% compared with that displayed on T2-weighted images, with only minor artifacts related to susceptibility at the skull base. Measured regional cerebral blood volume (rCBV) reduction closely correlated to lesion size on initial DWI and to final clinical outcome (P = 0.006), consistent with results previously reported for 2D perfusion methods. Mismatches between DWI and perfusion imaging characterized the total extent of tissue at risk, and the contrast timing correlated with the amount of collateral circulation as shown on DSA. In conclusion, 3D imaging using the PRESTO technique permits high-quality perfusion imaging of the entire brain.