Abstract: Objective To evaluate the feasibility and accuracy of automated apparentdiffusion coefficient imaging guided frameless stereotactic biopsy of intracranial lesions. Methods From January to July in 2015, a consecutive series of 16 patients with intracranial lesionsdiagnosed by biopsy were prospectively enrolled. The cohort consisted of 9 males and 7 females with a mean age of 46 years (ranged from 9 to 78 years). All patients had undergone apparentdiffusion coefficient (ADC) imaging-guided biopsy. Results ADC imaging-guided biopsy was successfully implemented in all cases. It took 2 minutes 7 seconds for single patient and 1 minute 53 seconds for single target todelineate targets using ADC imaging. All cases got a clear pathologicaldiagnosis with thediagnosis positive rate of 100%. Intracerebral hematoma and worse neurologicaldeficits was not found after surgery. Conclusion ADC imaging-guided frameless stereotactic biopsy of intracranial lesions has a high pathologicaldiagnosis positive rate. The target can be selected efficiently according to the ADC imaging of the lesion with high accuracy and safety.