To determine the relationship between reductions in the apparent diffusion coefficient of water (ADC) and in cerebral blood flow (CBF) during focal ischemia, we used diffusion-weighted magnetic resonance (D-MR) imaging and autoradiographic CBF analysis to examine rats subjected to 30 or 90 min of permanent middle cerebral artery (MCA) occlusion. In the 30-min occlusion group (n = l0), the area with substantially reduced ADC (15% or more below the contralateral level [ADCJ 15 ]) corresponded best to the area with CBF below 25 ml/lOO g/min and was significantly smaller than the area with CBF below 50 m1/100 g/min (CBF 50 ), a level associated with reduced protein synthesis and delayed necrosis (40 ± 13% versus 74 ± 8% of the ischemic hemisphere; P < 0.OOOl). In the 90-min occlusion group (n = 6), the ADC 15 area corresponded best to the CBF 30 to CBF 35 area and was again significantly smaller than the CBF 50 area (54 ± 13% versus 73 ± 20%, P < 0.05). Thus, the area of substantially reduced ADC at 30 and 90 min represents only 53% and 74%, respectively, of the tissue at risk for infarction. These findings indicate a potential limitation in using early D-MR imaging to predict stroke outcome.