The treatment of choice in Cushing's disease (CD) is surgical removal; however, most tumors are too small to be detected. The objective was to establish a method to achieve the complete removal of tumors on the basis of the results of high-resolution magnetic resonance imaging (MRI), inferior petrosal sinus sampling (IPSS), and a surgical resection technique using frozen biopsy. Eighteen patients who underwent transsphenoidal surgery from 2004 to 2010 were included. High-resolution MRI and IPSS, multiple-staged resection, and tumor tissue identification in frozen sections (surgical and histological identification, SHI) were performed. All patients achieved surgical remission, as confirmed by 24 h urinary free cortisol excretion tests. Visible microlesions were identified on the initial MRI in 11 patients (61%). The SHI findings agreed with the MRI findings in 10 of the 11 patients (90.9%) and with IPSS lateralization in 6 of the 11 patients (54.5%). In the 7 patients whose lesions were not visible on the initial MRI, only 1 (14.3%) showed an agreement between IPSS and SHI. In 3 of the 7 patients, the microlesions were identified by additional MRI. The rate of concordance with SHI was 77.8% for the overall MRI and 38.9% for IPSS. High-resolution MRI is better than IPSS for localizing corticotroph adenomas. In patients with lesions not visible on the initial MRI, additional MRI should be performed using a different protocol. Although high-resolution MRI is better for localizing tumors, SHI remains an important approach for removing the tumors completely.