Conventional quantitative assessments of myocardial perfusion analyze the temporal relation between the arterial input function and the myocardial signal intensity curves, thereby neglecting the important spatial relation between the myocardial signal intensity curves. The new method presented in this article enables characterization of sub-endocardial to subepicardial gradients in myocardial perfusion based on a two dimensional, "gradientogram'' representation, which displays the evolution of the transmural gradient in myocardial contrast uptake over time in all circumferential positions of the acquired images. Moreover, based on segmentation in these gradientograms, several new measurements that characterize transmural myocardial perfusion distribution over time are defined. In application to clinical image data, the new two-dimensional representations, as well as the newly defined measurements revealed a clear distinction between normal perfusion and inducible ischaemia. Thus, the new measurements may serve as diagnostic markers for the detection and characterization of epicardial coronary and microvascular disease. Magn Reson Med 66:1477-1487, 2011. (C) 2011 Wiley Periodicals, Inc.