To develop a less-stressful and simple method for measurement of the cerebral metabolic rate of oxygen (CMRO 2 ) in small animals, the steady-state method was applied to injectable 15 O 2 -PET ( 15 O 2 -positron emission tomography) using hemoglobin-containing vesicles ( 15 O 2 -HbV). Ten normal rats and 10 with middle cerebral arterial occlusion (MCAO) were studied using a small animal PET scanner. A series of 15 O-PET scans with C 15 O-labeled HbV, H 2 15 O, and 15 O 2 -HbV were performed with 10 to 15 minutes intervals to measure cerebral blood volume (CBV), cerebral blood flow (CBF), and CMRO 2 . Positron emission tomography scans were started with a tracer injection using a multiprogramming syringe pump, which provides a slowly increasing injection volume to achieve steady-state radioactivity for H 2 15 O and 15 O 2 -HbV scans. The radioactivity concentration of 15 O rapidly achieved equilibrium in the blood and whole brain at about 2 minutes after H 2 15 O and 15 O 2 -HbV administration, which was stable during the scans. The whole brain mean values of CBF, CBV, and CMRO 2 were 54.3 ± 2.0 mL per 100 g per minute, 4.9 ± 0.4 mL/100 g, and 2.8 ± 0.2 lmoL per g per minute (6.2±0.4 mL per 100 g per minute) in the normal rats, respectively. In the MCAO model rats, all hemodynamic parameters of the infarction area on the occlusion side significantly decreased. The steady-state method with 15 O-labeled HbV is simple and useful to analyze hemodynamic changes in studies with model animals.