CONCLUSIONS The determination of aniseikonia tolerance may aid in developing optimal treatment plans for cataract surgery, refractive surgery, and refractive correction with glasses and contact lenses. OBJECTIVE This study aimed to measure aniseikonia tolerance. METHODS We included 33 patients (mean age ± standard deviation, 28.9 ± 6.4 years; male/female, 12:21) with anisometropia ≤1.0 D and best spectacle-corrected visual acuity of 20/20 or more in both eyes, with no ophthalmologic disease other than refractive errors and no history of ocular surgery. The exclusion criteria were anisometropia >1.0 D, axial length difference >0.5 mm, corneal refractive power difference >0.5 D, astigmatism >3.0 D, stereoacuity threshold >100 arcsec according to the Titmus Stereo Test, and >0% aniseikonia according to the New Aniseikonia Test. Aniseikonia tolerance was assessed using Eyemark Hello, a haploscope using gaze detection technology. Although the optotype of one eye was enlarged or reduced at a speed of 2%/s, the patients were instructed to press a button on the controller to indicate blurring, flickering, and diplopia. The value at which the patient responded was considered the aniseikonia tolerance value and assessed thrice per eye, five times if the values were highly variable, and then averaged. RESULTS The mean aniseikonia tolerance was approximately 3%; the median value was approximately 2% (range, 1.0 to 11.5%; dominant eye, 3.3 ± 2.6%; nondominant eye, 2.9 ± 1.8%). No significant difference in aniseikonia tolerance between the dominant and nondominant eyes was observed for the enlarged optotypes. No case showed changes in the ocular alignment before discomfort occurred. No significant correlation was observed between aniseikonia tolerance and anisometropia, axial length difference, corneal power difference, and ocular deviation. CONCLUSIONS Aniseikonia should be maintained at <2% for a comfortable visual environment. Aniseikonia tolerance may be an important indicator for cataract surgery, refractive surgery, and spectacle correction.