Atrophy of central white matter is related to irreversible clinical disability in multiple sclerosis (MS) and ventricular enlargement may be a sensitive marker of this tissue loss. Therapeutic trials in MS have provided MRI data for investigation of cerebral atrophy in MS. These studies use almost exclusively two-dimensional (2-D) images, which may be limited in the assessment of three-dimensional (3-D) structures. We used 3-D MRI data to estimate ventricular volumes in 40 patients with MS and 10 healthy controls, to look at associations with clinical disability and the stage of the disease. We then compared simple linear measures of ventricular size from conventional 2-D images, with 3-D volume estimates to establish the best available linear indices of ventricular volume. Mean ventricular volumes were increased in the patients and significantly larger in the more disabled patients. The estimated volume of the third ventricle obtained from 3-D MRI showed the strongest association with the clinical stage of the disease, duration of symptoms and levels of disability. Finally, we confirmed that in patients with MS accurate data on ventricular size can be obtained from 2-D images by two simple and convenient linear measures, the width of the third ventricle and of the anterior horn of the lateral ventricle.