Endothelial vesicles on the luminal front of retinal and iridial vessels were studied by perfusion fixation after intravenous injection of peroxidase, and by perfusion fixation of glutaraldehyde with tannic acid or ruthenium red. Many luminal vesicles contained peroxidase reaction product after intravenous injection of peroxidase, but were virtually empty after perfusion fixation. Both tannic acid and ruthenium red delineated the majority of luminal vesicles, singular or in clusters, surface-associated or“free”in the cytoplasm. Only a few free vesicles were unlabeled by the dye. No transendothelial channels were seen. The results suggest that most of the vesicles on the luminal front of endothelial cells are continuous with the cell surface at the time of fixation. The function of these vesicles is not fully understood; some appear to be involved in endocytosis of macromolecules from the vascular lumen.