Reflection imaging of China ink‐perfused brain vasculature using confocal laser‐scanning microscopy after clarification of brain tissue by the Spalteholz method

• Published in: Journal of anatomy Vol. 230; no. 4; pp. 601 - 606
• Main Authors: Gutierre, R. C., Vannucci Campos, D., Mortara, R. A., Coppi, A. A., Arida, R. M.
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01-04-2017; John Wiley and Sons Inc
• Subjects: Animals; brain; Brain - blood supply; Brain - physiology; Brain Chemistry - physiology; Cerebrovascular Circulation - physiology; clarification; confocal microscopy; Ink; Laboratory animals; Lasers; Male; Methods; Microscopy; Microscopy, Confocal - methods; Rats; Rats, Wistar; Staining and Labeling - methods; stereology; vessels; stereology; clarification; brain; confocal microscopy; vessels
• ISSN: 0021-8782; 1469-7580
• DOI: 10.1111/joa.12578

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.