Fluorescence labeling of the capillary network in rat brains

To assess the reliability of fluorescence methods for a quantitative staining of brain capillaries, three different immunohistochemical fluorescent markers were used in the rat brain. Staining of the basement membrane by antibodies directed against fibronectin was compared, in the same brain section, with simultaneous staining of the vascular endothelium constituents nonmuscle myosin or von Willebrand factor (factor VIII). These stainings all resulted in identical patterns, which demonstrates their suitability for capillary staining in the brain. It has been claimed that fixation of the tissue results in the appearance of spurious capillary spots. Such a fixation artifact could be excluded using nonmuscle myosin staining. These results validate the methods of quantitative fluorescent microscopical staining of capillary morphology in the brain and therefore support our concept of a continuous perfusion of all capillaries in the brains of conscious rats.