Figure 1

The digital images were acquired at a single wavelength (ex 430 nm; em 510 nm). The recorded image stacks were analyzed for flow velocity (B) and recombined into a composite time-series image (C). A,B) A line of selectable orientation and width was drawn along the vessel axis. The distance-time plane (B) provided a longitudinal view over the selected length of the vessel. Cells or particles were tracked through multiple planes of the stack permitting a visual correlation in each plane. The white object (arrow) represents a fluorescent particle; the slope of the diagonal line represents the velocity of the particle in the flow stream. Note the different slope of the background speckle pattern--an observation suggesting the particle is near to, or interacting with, the vessel wall. A,C) The image stack was digitally recombined and pseudocolored for presentation as a time-series image (C). The region within the vessel demonstrating no detectable fluorescence (arrow) was defined as an intravascular pillar.