Objective To analyze the clinical diagnostic value of high resolution computed tomography (HRCT) and magnetic resonance imaging (MRI) in of temporal bone cholesteatoma. Methods A total of 107 patients (125 ears) with suspected cholesteatoma were recruited from September 2017 to March 2019, including 62 males and 45 females, aged from 3 to 79 years, with an average age of (40.14±18.8) years. Preoperative HRCT and MRI scans of temporal bone were performed 3 days to 2 weeks before operation. The results of the imaging scans were assessed by the radiologist and otological surgeon independently as suspected cholesteatoma and non-cholesteatoma. Intraoperative findings were recorded as cholesteatoma and non-cholesteatomatous tissues, which were confirmed by histopathology. Results The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of HRCT for cholesteatoma were 68.8%, 39.6%, 64.6% and 44.2% respectively, while those of diffusion-weighted MRI (MRI-DWI) were 96.1%, 91.7%, 94.9% and 93.6%. Comparing the results of primary and secondary operations, the accuracy of MRI-DWI in the diagnosis of recurrent cholesteatoma was significantly higher than HRCT (P<0.05). Conclusion The accuracy of MRI-DWI in the diagnosis of cholesteatoma is significantly higher than that of HRCT, especially in the diagnosis of recurrent cases.