The objective was to assess visibility and anatomy of the petromastoid canal in high-resolution CT. Computed tomography images of 188 patients were reviewed for delineation of the petromastoid canal. This bony canal connects the mastoid antrum with the cranial cavity and houses the subarcuate artery and vein. The diameter, obtained in the middle portion of the canal, was compared with the diameter of the vestibular and cochlear aqueduct in all patients, and absolute values measured in 20 cases. Collimation was 1 mm in 164 and 2 mm in 24 examinations. Additionally, temporal bone of a cadaver was imaged and microdissected. The petromastoid canal was identified bilaterally in all 164 scans that were obtained with a slice thickness of 1 mm. In 5 of the 24 patients imaged with a collimation of 2 mm, the canal was not visible, most probably due to partial-volume effects. The petromastoid canal had the same diameter as the cochlear aqueduct in 42/44 (right/left), exceeded it in 66/61 and was smaller in 75/78 cases. In comparison to the vestibular aqueduct it had an equal diameter in 38/41 (right/left), exceeded it in 63/61, and was rated as smaller in 82/81 temporal bones. Diameters for the canals were: petromastoid canal 0.51+/-0.04 mm; cochlear aqueduct 0.57+/-0.03; and vestibular aqueduct 0.63+/-0.06 mm. Microdissection of the specimen revealed the entire course of the canal and demonstrated a similar appearance of the structure as in the images. The petromastoid canal can easily be identified on high-resolution, thin-slice CT images. Knowledge of the anatomy of this bony canal prevents misinterpretation as pathological structure, such as fracture line, which might occur if this structure is not known.