Background: This study aimed to investigate the value of high field-strength intraoperative magnetic resonance imaging (iMRI)-guided stereotactic biopsy in the surgery of intracranial space-occupying lesions. Methods: A total of 87 patients who underwent stereotactic biopsy of intracranial lesions in the Peking University International Hospital from March 2016 to August 2018 were retrospectively surveyed; among these, 50 patients underwent MRI-guided stereotactic biopsy using the Leksell frame (iMRI group) and 37 cases received traditional stereotactic biopsy using the Leksell frame (control group). The accuracy rates and complications of the two groups were compared. Results: A 100% positive diagnosis was observed in all cases (n = 50) in the iMRI group. In 4 cases, the biopsy site was clearly found to have deviated from the target point, and the biopsy was performed again. The control group had 33 cases (86.5%) with positive diagnosis. No severe complications like neural functional deficit were observed in the iMRI group, while two patients developed bleeding at the puncture site (1 case receiving surgery to remove the hematoma) in the control group. There were no deaths in either group. Conclusion: iMRI-assisted stereotactic biopsy can confirm the target position and adjust the puncture path in real time. Compared to the traditional stereotactic biopsy technique, the iMRI method has a higher positive diagnostic rate, though surgical trauma and complications have no significant difference. © 2019 The Author(s).