This cadaveric anatomical study aimed to explore precise morphometric measurements of the eustachian tube (ET) and adjacent structures in the middle cranial fossa, focusing on identifying reliable surgical landmarks when traditional markers are compromised due to tumors or trauma. Twenty-two temporal bones from 11 adult cadavers (mean age: 75.70 ± 13.75 yr, range: 40-90 yr; sex: 5 females and 6 males) were dissected bilaterally. Surgical tools, including an operation microscope, endoscope, and digital caliper, were used for meticulous measurements. Parameters such as ET dimensions, distances between key points, and relevant angles were quantified, ensuring precise anatomical data. ET width at the foramen spinosum (FS) level, the midline level, and the eustachian orifice level were measured as 2.18 ± 0.68, 2.42 ± 0.70, and 2.30 ± 0.74 mm, respectively. The distances from the zygomatic root (ZR) to FS, ET, superior semicircular canal (SSC), and internal carotid artery (ICA) were 29.61 ± 2.56, 23.28 ± 2.61, 26.53 ± 2.56, and 32.61 ± 3.69 mm, respectively. The angles between SSC-ZR-ICA and FS-ZR-ICA were measured as 36.57 ± 10.32 and 13.63 ± 3.72 degrees, respectively. No statistical difference was found between right-left or male-female measurements ( p > 0.05). The present study offers invaluable insights for neurotological surgeons performing middle fossa approaches. ET and ZR may serve as crucial reference points, enhancing surgical orientation and minimizing risks during complex procedures. These precise anatomical data may empower surgeons, ensuring safer and more confident middle cranial fossa operations, even in challenging clinical scenarios.