The use of immunofluorescent immunohistochemical methods has been instrumental to characterize the distribution and extents of colocalization of molecules located in various cellular compartments. Such information has been pivotal to formulate hypothesis about their roles in physiological and pathological conditions.In the brain, astrocytes interact with endothelial cells to form the blood-brain barrier (BBB). Astrocytes regulate features of the BBB through the tips of their processes, called astrocyte endfeet, which surround and contact brain microvessels. Any disruption in astrocyte endfeet can have dramatic consequences on the integrity of the BBB, with consequent development of brain disorders.Here we describe a method to characterize the integrity of astrocyte endfeet around blood vessels in an animal model of major depressive disorder (MDD) using confocal microscopy, Adobe Photoshop, and the AutoQuantX3 software.
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