The objective of inter-subject registration of three-dimensional volumetric brain scans is to reduce the anatomical variability between the images scanned from different individuals. This is a necessary step in many different applications such as voxelwise group analysis of imaging data obtained from different individuals. In this paper, the ability of three different image registration algorithms in reducing inter-subject anatomical variability is quantitatively compared using a set of common high-resolution volumetric magnetic resonance imaging scans from 17 subjects. The algorithms are from the automatic image registration (AIR; version 5), the statistical parametric mapping (SPM99), and the automatic registration toolbox (ART) packages. The latter includes the implementation of a non-linear image registration algorithm, details of which are presented in this paper. The accuracy of registration is quantified in terms of two independent measures: (1) post-registration spatial dispersion of sets of homologous landmarks manually identified on images before or after registration; and (2) voxelwise image standard deviation maps computed within the set of images registered by each algorithm. Both measures showed that the ART algorithm is clearly superior to both AIR and SPM99 in reducing inter-subject anatomical variability. The spatial dispersion measure was found to be more sensitive when the landmarks were placed after image registration. The standard deviation measure was found sensitive to intensity normalization or the method of image interpolation.