title: Reflection imaging of China ink-perfused brain vasculature using confocal laser-scanning microscopy after clarification of brain tissue by the Spalteholz method.
abstract: Confocal laser-scanning microscopy is a useful tool for visualizing neurons and glia in transparent preparations of brain tissue from laboratory animals. Currently, imaging capillaries and venules in transparent brain tissues requires the use of fluorescent proteins. Here, we show that vessels can be imaged by confocal laser-scanning microscopy in transparent cortical, hippocampal and cerebellar preparations after clarification of China ink-injected specimens by the Spalteholz method. This method may be suitable for global, three-dimensional, quantitative analyses of vessels, including stereological estimations of total volume and length and of surface area of vessels, which constitute indirect approaches to investigate angiogenesis.
authors.affiliation: Department of Neurology and Neurosurgery, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.|Department of Physiology, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, Braz.
authors.forename: R C
authors.initials: RC
authors.lastname: Gutierre
authors.affiliation: Department of Neurology and Neurosurgery, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.|Department of Physiology, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, Braz.
authors.forename: D
authors.initials: D
authors.lastname: Vannucci Campos
authors.affiliation: Department of Microbiology, Immunology and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.
authors.forename: R A
authors.initials: RA
authors.lastname: Mortara
authors.affiliation: Faculty of Health and Medical Sciences, School of Veterinary Medicine, University of Surrey, Guildford, Surrey, UK.
authors.forename: A A
authors.initials: AA
authors.lastname: Coppi
authors.affiliation: Department of Neurology and Neurosurgery, Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil.
authors.forename: R M
authors.initials: RM
authors.lastname: Arida
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply* / Q000502:physiology; D001923:Brain Chemistry / Q000502:physiology; D002560:Cerebrovascular Circulation / Q000502:physiology; D007281:Ink; D008297:Male; D018613:Microscopy, Confocal / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D013194:Staining and Labeling / Q000379:methods*
keywords: brain; clarification; confocal microscopy; stereology; vessels