Glyoxal Fixation Allows Constant Staining of Brain Vessels, Pericytes, and Blood-Brain Barrier Proteins

Abstract Brain vascular staining has great importance in understanding cerebrovascular pathologies. Paraformaldehyde 4% is considered the gold standard fixation technique for immunohistochemistry and it revolutionized the examination of proteins in fixed tissues. However, this fixation technique produces inconsistent immunohistochemical staining results due to antigen masking. Here we test a new fixation protocol using 3% glyoxal and demonstrate that this method provides overall better staining of the brain vasculature, pericytes, and tight junction proteins than 4% paraformaldehyde. Using this new fixation technique will provide more detailed information about vascular protein expressions, their distributions, and colocalizations with other proteins at the molecular level in the brain vasculature.