ObjectiveTo determine whether 3D cone-beam computed tomography (CBCT) guidance allows safe and accurate biopsy of suspected small renal masses (SRM), especially in hard-to-reach anatomical locations.Materials and methodsCBCT guidance was used to perform 41 stereotactic biopsy procedures of lesions that were inaccessible for ultrasound guidance or CT guidance. In CBCT guidance, a 3D-volume data set is acquired by rotating a C-arm flat-panel detector angiosystem around the patient. In the data set, a needle trajectory is determined and, after co-registration, a fusion image is created from fluoroscopy and a slice from the data set, enabling the needle to be positioned in real time.ResultsOf the 41 lesions, 22 were malignant, 17 were benign, and 2 were nondiagnostic. The two nondiagnostic lesions proved to be renal cell carcinoma. There was no growth during follow-up imaging of the benign lesions (mean 29 months). This resulted in a sensitivity, specificity, PPV, NPV, and accuracy of 91.7, 100, 100, 89.5, and 95.1%, respectively. Mean dose-area product value was 44.0 Gy·cm2 (range 16.5–126.5). There was one minor bleeding complication.ConclusionWith CBCT guidance, safe and accurate biopsy of a suspected SRM is feasible, especially in hard-to-reach locations of the kidney.Key Points• Cone-beam computed tomography has potential advantages over conventional CT for interventional procedures.• CBCT guidance incorporates 3D CBCT data, fluoroscopy, and guidance software.• In hard-to-reach renal masses, CBCT guidance offers an alternative biopsy method.• CBCT guidance offers good outcome and safety and has potential clinical significance.