Quantitative MRI assessment of leukoencephalopathy is difficult because the MRI properties of leukoencephalopathy significantly overlap those of normal tissue. This report describes the use of an automated procedure for longitudinal measurement of tissue volume and relaxation times to quantify leukoencephalopathy. Images derived by using this procedure in patients undergoing therapy for acute lymphoblastic leukemia (ALL) are presented. Five examinations from each of five volunteers (25 examinations) were used to test the reproducibility of quantitated baseline and subsequent, normal-appearing images; the coefficients of variation were less than 2% for gray and white matter. Regions of leukoencephalopathy in patients were assessed by comparison with manual segmentation. Two radiologists manually segmented images from 15 randomly chosen MRI examinations that exhibited leukoencephalopathy. Kappa analyses showed that the two radiologists' interpretations were concordant (κ = 0.70) and that each radiologist's interpretations agreed with the results of the automated procedure (κ = 0.57 and 0.55).The clinical application of this method was illustrated by analysis of images from sequential MR examinations of two patients who developed leukoencephalopathy during treatment for ALL. The ultimate goal is to use these quantitative MR imaging measures to better understand therapy-induced neurotoxicity, which can be limited or even reversed with some combination of therapy adjustments and pharmacological and neurobehavioral interventions. Magn Reson Med 47:912–920, 2002. © 2002 Wiley-Liss, Inc.