Hubei Provincial Clinical Research Center for Diabetes and Metabolic Disorder 1 article published in JoVE Neuroscience Free-floating Immunostaining of Mouse Brains Longlong Tu1, Nan Zhang1,2,3, Kristine M Conde1, Jonathan C Bean1, Chunmei Wang1, Yong Xu1,4 1USDA/ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 2Department of Endocrinology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 3Hubei Provincial Clinical Research Center for Diabetes and Metabolic Disorder, 4Department of Molecular and Cellular Biology, Baylor College of Medicine This protocol describes an efficient and reproducible approach for mouse brain histological studies, including perfusion, brain sectioning, free-floating immunostaining, tissue mounting, and imaging.