Fluid accumulation within the tympanic bulla is an important diagnostic indicator of canine otitis media although its identification can be a challenge using currently available imaging techniques. The purpose of this study was to compare radiography, computed tomography (CT) and magnetic resonance imaging (MRI) in the identification of fluid accumulation within canine tympanic bulla. Unilateral tympanic bulla in 10 beagles were experimentally filled with blood or saline. Quantitative analysis of CT images were obtained by using Hounsfield unit (HU). MR signal intensity was obtained by using region of interesting (ROI) and compared with those of gray matter. On the CT image, the presence of blood or saline produced a fluid opacity occupying the tympanic bulla. On the MR image, the appearance of blood in the tympanic bulla was isointense in T1-weighted images and hyperintense in T2-weighted images. However, the appearance of saline in the tympanic bulla was hypointense in T1-weighted images and hyperintense in T2-weighted images. This study suggest that CT and MR imaging are useful methods for detection and differentiation of fluid in canine tympanic bulla.