PurposeTo evaluate the effect of freezing and thawing on the biomechanical properties of ex-vivo porcine ocular tissue. MethodsThirty-six porcine eyes (18 pairs) were obtained fresh from a local abattoir and split into two groups of nine pairs to study the effect of storage at −20 °C and −80 °C. A randomly-selected eye from each pair (Control Group, CG) was tested fresh while the fellow eyes were frozen for 14 days, either at −20 °C and −80 °C (Frozen Group, FG) before thawing and testing. Seventy-two strips were extracted from the corneas and scleras of eye globes and subjected to uniaxial tension tests under loads up to 1.0 N. Following five preconditioning cycles, the load and elongation data obtained experimentally were analysed to derive the tissue’s stress-strain and tangent modulus-strain behaviour. ResultsCorneal tissue subjected to freezing at −20 °C exhibited significant increases in tangent modulus (mechanical stiffness) by 13 ± 17% (p = 0.003) at 1% strain and 14 ± 12% (p < 0.001) at 2% strain. In contrast, the increases in corneal stiffness at −80 °C were insignificant (6 ± 14%, p = 0.099 at 1% strain, 6 ± 15%, p = 0.091 at 2% strain). The corresponding increases in tangent modulus in the sclera were all insignificant (for −20 °C: 4 ± 14%, p = 0.265 at 1% strain, 3 ± 9%, p = 0.186 at 2% strain; for −80 °C: 3 ± 18%, p = 0.537 at 1% strain and 3 ± 18%, p = 0.491 at 2% strain). ConclusionsThe study provided evidence that freezing and thawing led to insignificant changes in ocular tissue stiffness except in corneal tissue that was frozen at −20 °C.