Tympanokeratomas (cholesteatomas) are destructive keratinic masses of the middle ear that are often treated surgically. In humans, nonechoplanar (non-EP) diffusion-weighted (DW) MRI is efficient in diagnosing tympanokeratomas and differentiating recurrent tympanokeratomas from granulation or fibrous tissue after surgery. The objectives of this study were to (1) determine the characteristics of non-EP DWI of histologically or cytologically confirmed canine tympanokeratomas, (2) determine the performance of non-EP DW images, apparent diffusion coefficient (ADC) map and ADC values in discriminating between tympanokeratomas and other causes of otitis media in dogs, and (3) find an optimal ADC cut-off value. Medical records of dogs diagnosed on MRI with otitis media in two veterinary hospitals were retrospectively reviewed. Sixty-two ears with a cytological or pathological diagnosis of either tympanokeratoma (36/62) or nontympanokeratoma otitis media (26/62) were selected. Diffusion-weighted images had an accuracy of 77.4%, a sensitivity of 94.4%, and a specificity of 53.8% for the detection of tympanokeratoma. The ADC value of middle ear content ranged from 517 to 1355.10-6mm2/s in tympanokeratoma and from 454 to 2447×10-6mm2/s in nontympanokeratoma otitis media. An ADC value below 916×10-6mm2/s interpreted as a tympanokeratoma yielded an accuracy of 78.7%, a sensitivity of 92.6%, and a specificity of 60%. In the study population, combined qualitative analysis of DW images and ADC maps was the most effective method for diagnosing tympanokeratomas with an accuracy of 80.9%, a sensitivity of 92.6%, and a specificity of 65%.