Using theoretical models for the flow of fluourescein dye in retinal arteries and veins, we have determined the effects of optical absorption in blood of the incident excitation light and the emitted fluorescent light on the time course of measured fluorescence intensity, I(t). Our results indicate that I(t) curves recorded from arteries adequately represent the mean time course of the fluorescein concentration (C(t)), when either a circular or rectangular light-collecting aperture is used. I(t) curves recorded from veins adequately represent C(t), but only when a circular aperture of approximately the same diameter as that of the vessel is used. A two-point fluorophotometer, which provides simultaneous, on-line measurements of arterial and venous I(t) curves is described. Typical recordings obtained with the instrument are shown and the method employed to analyze the curves quantitatively is described in detail. This method, which consists of fitting the first passage of the fluorescence intensity curve with a log-normal function, provides results that are more accurate than those obtained using the standard exponential extrapolation method.