Diffusion-weighted magnetic resonance imaging (DW-MRI) provides a unique form of MR contrast that enables the diffusional motion of water molecules to be quantitatively measured. As a consequence, DW-MRI provides information about the size, shape, integrity, and orientation of brain structures. Pathological processes able to alter tissue integrity by removing or modifying some of the structural barriers that normally restrict water molecular motion in biological tissues cause changes in water diffusion characteristics, which can be measured in-vivo using DW-MRI. Although DW-MRI has been shown to be of great clinical utility in the assessment of patients with cerebral ischemia, it is also increasingly being used to quantify in-vivo the extent and severity of multiple sclerosis (MS) pathology. The pathological elements of MS have the potential to alter the permeability or geometry of structural barriers to water molecular motion in the brain, optic nerve and spinal cord. The present review outlines the major contributions given by DW-MRI for the quantification of MS-related damage and for the understanding of MS pathophysiology.