Purpose: The present study aimed to assess the accuracies of arterial stimulation with simultaneous venous sampling(ASVS) in preoperative localization of insulinomas. Materials and Methods: A cohort consisting of 6 males and 14 females(median age, 48.5y;range, 28–62y) with pathologically proven insulinomas were included in this study. Selective angiographies were performed with the superior mesenteric artery(SMA), gastroduodenal artery(GDA), proximal splenic artery, and midsplenic artery in all individuals. Then ASVS procedures were followed after angiographies for these arteries. Clinical characteristics of the patient and the tumor number, location, and size were recorded. The accuracy of preoperative localization of insulinomas was tested. Results: A total of 22 tumors were identified by histopathological diagnosis. The mean size of the tumor was 1.40±0.60 cm. Five tumors were in the head/neck region and 17 in the body/tail region. ASVS accurately localized 17/20(85%) cases with only biochemical data and 19/20(95%) cases with biochemical data and angiography images. Variant pancreatic arterial anatomy was revealed in 2 false cases with inferior pancreatic artery replaced by the superior mesenteric artery. Conclusion: ASVS was highly accurate in localizing insulinomas and should be performed in most of the patients with suspected insulinomas before the operation.