Endothelial damage is considered to be an initial change in the atherosclerotic process. However, it has been difficult to detect this initial change in vivo. We established a modified En face immunostaining method that enabled us to obtain clear images of the entire endothelial surface, including at arterial bifurcations, and to quantitate the number of cells of interest in the endothelium. Using this method, we found that treatment with an atherogenic factor, albumin-derived advanced glycosylation end products, for only 2 weeks caused a significant increase in the number of proliferating cell nuclear antigen-positive endothelial cells and the number of macrophages adhering to the endothelium, suggesting that these changes might be relevant to the early events of endothelial dysfunction. In conclusion, the present modified En face immunostaining method may be a promising tool for understanding the pathophysiology of atherosclerosis. (C) 2003 Elsevier Inc. All rights reserved.