Anisometropic amblyopia is decreased visual acuity in one eye, and treatment consists of wearing complete corrective spectacles. Aniseikonia occurs with complete correction of anisometropia using spectacles. Aniseikonia has been ignored when treating pediatric anisometropic amblyopia because of the prevailing belief that anisometropic symptoms are suppressed by adaptation. However, the conventional direct comparison method for evaluating aniseikonia significantly underestimates the degree of aniseikonia. This study investigated whether the adaptation occurs due to long-term anisometropic amblyopia treatment in patients who have had successful amblyopia treatment using a spatial aniseikonia test with high accuracy and repeatability compared with the conventional direct comparison method. The amount of aniseikonia was not significantly different between the patients with successful amblyopia treatment and individuals with anisometropia without a history of amblyopia. In both groups, the aniseikonia per 1.00 D of anisometropia and the aniseikonia per 1.00 mm of aniso-axial length were comparable. The repeatability of the amount of aniseikonia using the spatial aniseikonia test did not differ significantly between the two groups, indicating a high degree of agreement. These findings suggest that aniseikonia is not adapted to amblyopia treatment and that aniseikonia increases as the difference between spherical equivalent and axial length increases.