Currently available clinical devices are unable to measure corneal biomechanics other than at the central region. Corneal stiffness (S), thickness, and radius of curvature was measured at the central cornea (primary fixation) and 3 mm from the temporal limbus (primary and nasal fixations). The corneal tangent modulus (E) of 25 healthy subjects was calculated from these data. After confirming normality, repeated measures analysis of variance (RMANOVA) revealed significant difference in S (F(2, 48) = 21.36, p < 0.001) at different corneal regions and direction of fixations. E also varied significantly at different corneal regions and direction of fixations (RMANOVA: F(2, 48) = 23.06, p < 0.001). A higher S and a lower E were observed at the temporal region compared with the corneal centre. Nasal fixation further increased S and E values compared with primary fixation. Due to the specific arrangement of corneal collagen fibrils, heterogeneity of corneal biomechanical properties is expected. In future clinical practice, localized corneal biomechanical alternation and measurement might assist corneal disease detection and post-surgery management. In addition, practitioners should be aware of the fixation effect on corneal biomechanical measurement.