BACKGROUND: A recent report suggests that 70%-80% Z-tenotomy of the superior oblique tendon is necessary to effectively treat A-pattern strabismus associated with over depression in adduction. To clarify the clinical effect, we compared the biomechanics of Z-tenotomy on the superior oblique tendon, superior rectus tendon, and isotropic latex material. METHODS: Fresh bovine superior oblique tendons were trimmed to 20 mm × 10 mm dimensions similar to human superior oblique tendon and clamped in a microtensile load cell under physiological conditions of temperature and humidity. Minimal preload was applied to avoid slackness. Tendons were elongated until failure following Z-tenotomies, made from opposite tendon margins, spaced 8 mm apart and each encompassing 0%, 20%, 40%, 50%, 60%, or 80% tendon width. Digitally sampled failure force was monitored using a precision strain gauge. Control experiments were performed in similar-sized specimens of bovine superior rectus tendon and isotropic latex. RESULTS: Progressively increasing Z-tenotomy of latex caused a linearly graded reduction in force. In contrast, Z-tenotomy of up to 50% in superior oblique and superior rectus tendons caused nonlinear reduction in force transmission that reached a negligible value at 50% tenotomy and greater. CONCLUSIONS: Z-tenotomy up to 50% progressively reduces extraocular tendon force transmission, but Z-tenotomy of ≥50% is biomechanically equivalent in vitro to complete tenotomy.