AIM: To investigate the significance of ultrasound elastography for evaluating stiffness of the human lens nucleus in patients with anisometropia. METHODS: A total of 14 patients (28 eyes) with anisometropia were enrolled. The difference in refractive status between two eyes ≥-4.0 diopters (D) and the difference in axial length (AL) of the eyes was ≥3 mm. There were 5 males and 9 females with an average age of 62±3.3y. The test data of the long AL eye of each patient was included in group A (14 eyes), and test data of the eye with relative short AL was included in group B. Lens nuclear stiffness was measured with free-hand qualitative elastography by independent operators. Strain gray scale and color-coded elastography maps were recorded. In each case, three consecutive measurements were performed and strain ratio was used for statistical analysis. Photograph and sectional view of the lens were analyzed and archived by anterior segment image. RESULTS: In the long AL group, the strain rate in the nucleus of the lens was 0.16%±0.08%; in the short AL group, the strain rate in the nucleus of the lens was 0.54%±0.16%. Independent sample t-test analyses showed that the long AL group lens had a significantly smaller nuclear strain rate than the relatively short AL group, P<0.05. CONCLUSION: The relationship between human lens stiffness and different AL is demonstrated by ultrasound elastography. The long AL is associated with lower strain ratio and less resilience of the lens.