OBJECTIVE: The aim of this study was to compare the accuracy of Bolton analysis obtained from digital models scanned with the Ortho Insight three-dimensional (3D) laser scanner system to those obtained from cone-beam computed tomography (CBCT) images and traditional plaster models. METHODS: CBCT scans and plaster models were obtained from 50 patients. Plaster models were scanned using the Ortho Insight 3D laser scanner; Bolton ratios were calculated with its software. CBCT scans were imported and analyzed using AVIZO software. Plaster models were measured with a digital caliper. Data were analyzed with descriptive statistics and the intraclass correlation coefficient (ICC). RESULTS: Anterior and overall Bolton ratios obtained by the three different modalities exhibited excellent agreement (> 0.970). The mean differences between the scanned digital models and physical models and between the CBCT images and scanned digital models for overall Bolton ratios were 0.41 +/- 0.305% and 0.45 +/- 0.456%, respectively; for anterior Bolton ratios, 0.59 +/- 0.520% and 1.01 +/- 0.780%, respectively. ICC results showed that intraexaminer error reliability was generally excellent (> 0.858 for all three diagnostic modalities), with < 1.45% discrepancy in the Bolton analysis. CONCLUSIONS: Laser scanned digital models are highly accurate compared to physical models and CBCT scans for assessing the spatial relationships of dental arches for orthodontic diagnosis.