The objective of this study was to test for the presence of transmural gradients of various components of the coronary microvasculature of the canine left ventricle. In order to achieve study objectives, the heart and coronary circulation were fixed in a reproducible state of myocardial and vascular tone (diastolic cardiac arrest and maximal coronary vasodilation). Morphometric methods which treat the coronary microvasculature as anisotropically arranged structures were applied for quantitative structural analysis. Eight dog hearts were fixed with a glutaraldehyde-cacodylate-buffered fixative by retrograde perfusion of the aorta with the heart in diastolic arrest and with maximal coronary vasodilation. Tissue samples were taken from areas near to the anterior and posterior papillary muscles from the subendocardium, subepicardium, and intermediate transmural locations. Morphometric results showed a homogeneously arranged array of microvascular and myocardial components with no significant differences in any of the primary morphometric measurements, down to the ultrastructural level, in myocytes relative to transmural location. The results suggest that transmural differences in coronary blood flow are not due to transmural structural differences but rather are due to physiological regulatory mechanisms of coronary blood flow. Further, the results indicate that failure to correct for anisotropy of myocardial structures can lead to erroneous conclusions concerning the structural basis of function in the heart.