Purpose: To evaluate the effect of interocular axial length (AL) difference on outcomes of treatment for anisometropic amblyopia in comparison with normal participants. Methods: In this historical cohort study, 83 patients with anisometropic amblyopia were divided into two age groups, 70 children (mean, 7. 86 ± 1. 56 and range, 5–15 years) and 13 adults (mean, 26. 46 ± 10. 87 and range, 16–45 years). The control group consisted of 43 non-amblyopic children and 17 non-amblyopic adults. Treatment outcomes after a period of one year were defined as successful or unsuccessful when posttreatment amblyopic corrected distance visual acuity (CDVA) was reported as ≤0. 9 versus CDVA ≤ 0. 8, respectively. AL was measured using a Lenstar LS900 (Haag-Streit AG, Switzerland). Results: Fifty-nine patients showed satisfactory treatment outcomes (55 children and 4 adults), while unsuccessful treatment outcomes were observed in 24 patients (15 children and 9 adults). The mean of amblyopia treatment duration was 1. 24 ± 0. 76 years. The mean of interocular AL difference in all patients, control, successful and unsuccessful treatment outcome groups were 0. 49 ± 0. 70mm (range, 0. 00–3. 89 mm), 0. 12 ± 0. 07 mm (range, 0. 02–0. 41), 0. 33 ± 0. 23 mm (range, 0. 00–0. 99 mm), and 1. 81 ± 0. 80 mm (range, 1. 14–3. 89 mm), respectively. In both age groups, the mean of interocular AL difference in patients with unsuccessful treatment outcomes was greater than those with successful treatment outcomes and that of the control group (P < 0. 001). Conclusion: The results of this study suggest that the outcome of anisometropic amblyopia treatment may depend on the interocular AL difference.