Peripheral arterial occlusive disease (PAOD) is a leading cause of mortality and morbidity in the western world. The development of noninvasive methods for assessment and comparison of the efficacy of novel therapies in animal models is of great importance. Methods: Hindlimb ischemia was induced in nude mice by ligation and excision of the left femoral artery (n 5 5) or the left iliac artery (n 5 10). Assessment of limb perfusion was performed by small-animal PET analysis after intravenous injection of 13 Nammonia between 24 h and 30 d after surgery using the ratio of perfusion between the left limb (ischemic) and the right limb (control). Activity concentration per area unit was calculated in regions of interest placed on 1-mm-thick images for numeric calculations, and the iliac and the femoral models were compared. In addition, histopathologic studies were performed to assess the degree of necrosis (hematoxylin2eosin) and fibrosis (sirius red). Immunohistochemistry analyses for identification of arterioles (a-smooth muscle actin) and endothelium-capillaries-(Bandeiraea simplicifolia I [BS-I] lectin) were also performed. Results: Perfusion in both hindlimbs of control animals was similar (median of the left-to-right ratio 5 0.99). Twenty-four hours after ischemia, perfusion of the ischemic limb (% mean 6 SD) was 33.3 6 10.6 and 22.1 6 9.9 in the femoral and iliac models, respectively. Spontaneous recovery of perfusion in the hindlimb that underwent surgery was significantly lower in the iliac model at day 115 (73.2 6 15.5 vs. 51.9 6 11.3; P , 0.01). Fibrosis increased progressively until day 130, whereas muscle necrosis was maximal at day 17 with a moderate reduction by day 130. In accordance with this positive effect, there was a statistically significant increase in the area covered with smooth muscle-coated vessels (arterioles) at day 130 in comparison with day 7 (P , 0.05). In addition, a correlation between 13 N-ammonia uptake and the amount of necrosis (r 5 20.73; P 5 0.06) and fibrosis (r 5 20.67; P 5 0.05) at day 130 was found. Conclusion: 13 NAmmonia imaging allows semiquantitative evaluation of hindlimb perfusion in surgical mouse models of acute hindlimb ischemia. Although spontaneous perfusion recovery is observed in both models, the iliac model shows a substantially lower recovery and is hence better suited for assessment of new therapeutic strategies for acute hindlimb ischemic disease.