The guinea pig constitutes one of the most frequently used animal in otolaryngologic experimental research especially in the otologic research field. However, few has detailed on the anatomical study of this animal and the information available for temporal bone anatomy has been limited. In an attempt to be a help for further study and to reduce the trial and error, we investigated the temporal bone of both living and dead, and comparative anatomical study was done to discuss the similarity and dissimilarity between the human and the guinea pig. The authors dissected the temporal bone through both superior approach and inferior approach, and examined external auditory canal, tympanic membrane, ossicles, epitympanum, stapedius muscle, cochlea, Eustachian tube, semicircular canal, round window, and facial nerve under a surgical microscope. We noticed that the surgical approach of guinea pig is relatively easy and the temporal bone and ear structure of guinea pig are good models for the comparative study.