High resolution video fluorescein angiography using scanning laser ophthalmoscopy allows the assessment of retinal macro- and microcirculation. Data on the retinal macrocirculation were obtained from 221 healthy subjects. The data were derived from estimations of the arm-retina time, the arteriovenous passage time and mean arterial dye velocity, characterizing the passage of fluorescein to the eye, the mean arterial plasma velocity, and the arteriovenous passage through the entire vascular bed of one segment. Additionally, the transit of hypofluorescent segments in the capillary macular network were measured in 90 healthy subjects. These parameters provide a wide range of information for understanding the physiology of healthy and diseased eyes. Fundamental for all interpretations is the knowledge of the physiological variations. In the present study the inter- and intraindividual variability of retinal hemodynamics in healthy volunteers were assessed. The interindividual variation was 23.8% for the arm-retina time, 20.7% for the arteriovenous passage time, 23.7% for the mean arterial dye velocity, and 14.2% for the capillary flow velocity; the coefficient for variation, characterizing the intraindividual variation, was 26.6%, 15.6%, 16.7%, and 7.9%, respectively. The knowledge of the inter- and intraindividual variation of retinal blood flow indices allows for a priori power estimations for pathophysiologic and pharmacological studies.