AIM: To assess lens thickness measurements with anterior segment-optical coherence tomography (AS-OCT) in comparison with A-scan ultrasonography (A-scan US). METHODS: There were 218 adult subjects (218 eyes) aged 59.2±9.2y enrolled in this prospective cross-sectional study. Forty-three eyes had open angles and 175 eyes had narrow angles. Routine ophthalmic exam was performed and nuclear opacity was graded using the Lens Opacities Classification System III (LOCS III). Lens thickness was measured by AS-OCT (Visante OCT, Carl Zeiss Meditec, Dublin, CA, USA). The highest quality image was selected for each eye and lens thickness was calculated using ImageJ software. Lens thickness was also measured by A-scan US. RESULTS: Interclass correlations showed a value of 99.7% for intra-visit measurements and 95.3% for inter-visit measurements. The mean lens thickness measured by AS-OCT was not significantly different from that of A-scan US (4.861±0.404 vs 4.866±0.351 mm, P=0.74). Lens thickness values obtained from the two instruments were highly correlated overall (Pearson correlation coefficient=0.81, P<0.001), and in all LOCS III specific subgroups except in grade 5 of nuclear opacity. Bland-Altman analysis revealed a 95% limit of agreement from -0.45 to 0.46 mm. Lens thickness difference between the two instruments became smaller as the lens thickness increased and AS-OCT yielded smaller values than A-scan US in thicker lens (β=-0.29, P<0.001) CONCLUSION: AS-OCT-derived lens thickness measurement is valid and comparable to the results obtained by A-scan US. It can be used as a reliable noncontact method for measuring lens thickness in adults with or without significant cataract.