Objectives:The aim of this study is to evaluate the accuracy of linear measurements on cone-beam computed tomography (CBCT) images using three software programs and different voxel sizes.Methods:Ten human mandibles with 25 silica markers were scanned for 0.250-, 0.300-, and 0.400-mm voxels in the i-CAT New Generation (Imaging Sciences International, Hatfield, PA, USA). Thirty-five linear measurements were carried out by two examiners two times on the multiplanar reconstructions in the following software programs: XoranCat version 3.1.62 (Xoran Technologies, Ann Arbor, MI, USA), RadiAnt DICOM 2.2.9 Viewer (Medixant, Poznan- Poland) and InVesalius 3.0.0 (Centro de Tecnologia da Informação Renato Archer, Campinas, SP, Brazil). The physical measurements were made by another observer two times using a digital caliper on the macerated mandibles. ANOVA test was used to compare voxels and software programs. Pearson correlation and the Bland–atman tests were used to compare physical and virtual measurements and to evaluate the accuracy of the software programs, respectively (P < 0.05).Results:There was no statistically significant difference when the measurements were compared in acquisitions with different voxel sizes analyzed in the three software programs. There was also no difference when the measurements were compared between the software programs and the digital caliper. Excellent intra- and inter-observer reliability for the markers, physical measurements, and multiplanar reconstructions were found.Conclusion:Linear measurements in the XoranCat, Radiant, and InVesalius software programs are reliable and accurate compared with physical measurements. The different acquisition protocols using different voxel sizes did not influence the accuracy of linear measurements in CBCT images.