We developed a method for autoradiographic mapping of regional cerebral transit time (CTT) by simultaneously measuring local cerebral blood flow (LCBF) and local cerebral blood volume (LCBV). Previously described single-tracer techniques for determination of LCBF and LCBV were modified for dual-tracer, 99mTc and 14C, autoradiography and used to create digital images of LCBF and LCBV from the same brain sections in a series of normal rats. The images were aligned and ratio images (LCBV/LCBF) were then generated which reflected CTT. Regional cerebral transit time was found to vary significantly through-out the brain in a pattern only partially related to that of blood flow. Such CTT heterogeneity could cause errors in implementation of kinetic models which assume uniform or monovariant distributions of vascular transit time.