Velocity profiles were determined in rabbit mesenteric arterioles (diameter 17-32 /xm). A good spatial resolution was obtained by using the blood platelets as small and natural markers of flow, providing for the first time in vivo detailed, quantitative information about the shape of the velocity profiles in micro vessels. In some experiments red blood cell velocity profiles were recorded as well. Easy detection of the cells of interest could be achieved by labelling them selectively with a fluorescent dye and visualizing them by intravital fluorescence video microscopy, using flashed illumination. Pairs of flashes were given with a short, preset time interval between both flashes, yielding in one TV picture two images of the same cell displaced over a certain distance for the given time interval. Velocity and mean radial position of cells, flowing within an optical section around the median plane of the vessel, were determined. The shape of the velocity profiles of platelets and red blood cells was similar. The profiles were flattened as compared to a parabola, both in systole and diastole. Vessel diameter did not change measurably during the cardiac cycle. As an index of the degree of blunting of the profiles, the ratio of the maximal and mean velocity of the profile was used, which is 2 for a parabola and 1 for complete plug flow. The index ranged from 1.39 to 1.54 (median 1.50), and increased with vessel diameter. Calculations showed that the blunting of the profiles cannot be explained by an influence of the finite depth of the optical section. (Circulation Research 1986;59:505-514) knowledge of the velocity profile in microvessels, i.e., the velocity distribution over the cross-sectional area of these vessels, is important for several reasons. First, adequate description of blood flow through small vessels requires information about the velocity gradients or shear rates in the fluid.1 Second, transport of cellular components in the blood is determined by both their distribution over the cross-sectional area of the vessel and the velocity profile. For instance, knowledge of the distribution of blood platelets in microvessels 2 and their velocity profile allows the calculation of the rate of platelet delivery in hemostatic plug or thrombus formation in these vessels. Third, to estimate volume flow in microvessels photometric methods are widely used, 3 employing an empirical factor derived from a model in which a parabolic velocity profile is assumed. However, in this approach an error will be made if in vivo the velocity profiles are more flattened.In vitro studies in glass tubes on the velocity profiles of ghost cell suspensions 5 Until now, precise measurement of velocity profiles in small blood vessels has not been performed in vivo for technical reasons. Photometric methods cannot be used to determine a profile because the system does not provide a direct measure of the red blood cell velocity in the plane of sharp focus. 34 With high-speed cinematography, displacement of red blood cells can be followe...