Image-based vessel-by-vessel analysis for red blood cell and plasma dynamics with automatic segmentation

The aim of the present study was to test the hypothesis that vascular tones of cortical surface and parenchymal blood flow can be dissociated depending on the perturbation. To this end, a novel image-based analytical method for quantitatively measuring vessel diameters and flow dynamics was developed. The algorithm relies on the spatiotemporal coherence of the pixel intensity changes induced by the transit of the fluorescent signals measured using confocal laser scanning fluorescent microscopy in the rat cerebral cortex. A cocktail of fluorescently labeled red blood cell (RBC) and plasma agents was administered to simultaneously compare RBC and plasma dynamics in the same vascular networks. The time to fluorescent signal appearance and the width of the fluorescent signal were measured in each segment and compared between sodium nitroprusside-induced global and sensory stimulation-induced local perturbation conditions. We observed that infusion of sodium nitroprusside induced significant vasodilation in the surface artery, particularly in the small arteries (1.8-fold increase). Vasodilation induced by sensory stimulation was observed to depend on vessel size, but significant changes were only detected for the small arteries and veins. Measurements of the time to venous appearance revealed that appearance time was extended by sodium nitroprusside, but shortened during forepaw stimulation, relative to the control condition. Both perturbations provoked the largest changes between the small artery and vein segments, indicating that the changes in the appearance time originate from blood passage through parenchymal microcirculation. These findings support the hypothesis that cortical surface vascular tone and parenchymal blood flow are individually coordinated.