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Reflection imaging of China ink‐perfused brain vasculature using confocal laser‐scanning microscopy after clarification of brain tissue by the Spalteholz method
Author(s) -
Gutierre R. C.,
Vannucci Campos D.,
Mortara R. A.,
Coppi A. A.,
Arida R. M.
Publication year - 2017
Publication title -
journal of anatomy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 118
eISSN - 1469-7580
pISSN - 0021-8782
DOI - 10.1111/joa.12578
Subject(s) - confocal , confocal microscopy , microscopy , confocal laser scanning microscopy , laser scanning , hippocampal formation , confocal laser scanning microscope , biomedical engineering , pathology , laser microscopy , materials science , laser , biology , optics , medicine , neuroscience , microbiology and biotechnology , physics
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.