Objective: The study aimed to determine the prevalence, causes, and associations with amblyopia in a defined older population. Design: In a population-based study, 3654 persons 49 years of age or older from an area west of Sydney, Australia, underwent a detailed eye examination and history, including objective and subjective refraction, cover testing, and retinal and lens photography. Amblyopia was diagnosed in eyes with reduced best-corrected visual acuity in the absence of any other cause. Results: Amblyopia was diagnosed in 118 participants, or 3.2% of the population using a visual acuity criterion of 20/30 or less and 2.9% using a visual acuity criterion of 20/40 or less. Using a two-line visual acuity difference between the eyes, the amblyopia prevalence was 2.6% and 2.5%, respectively, for the above criteria. The underlying amblyogenic causes assessed were anisometropia (50%), strabismus (19%), mixed strabismus and anisometropia (27%), and visual deprivation (4%). The visual acuity of the amblyopic eye was 20/200 or worse (19%), 20/80 to 20/160 (19%), 20/40 to 20/63 (52%), and 20/30 (11%). No statistically significant associations were found between amblyopia and gender or eye affected. The most frequent pattern of strabismus was esotropia, whereas hypermetropia was the most frequent refractive error in amblyopic eyes. The mean age at diagnosis was earlier for strabismic and mixed amblyopia (7.4 years) than for anisometropic amblyopia (12.7 years). Conclusion: This study has provided prevalence and cases of amblyopia in an older population. Amblyopia is a frequent cause of lifelong unilateral visual impairment.