High-resolution Confocal Imaging of the Blood-brain Barrier: Imaging, 3D Reconstruction, and Quantification of Transcytosis

The overall goal of this method is to provide a microscopy-based protocol for high-resolution imaging and 3-dimensional reconstruction of the mouse blood-brain barrier.It combines immunofluorescence of free-floating sections with confocal microscopy to analyze subcellular organelles within the brain endothelial cells.This method add learns the preparation of brain free-floating sections, immunofluorescence staining, image acquisition and analysis parameters for high-resolution microscopy of the blood-brain barrier.The main advantage of the technique is that it allows for the quantification of subcellular organelles within cells of the neurovascular unit.Demonstrating the procedure will be Roberto Villase