Tissue Clearing and Light Sheet Microscopy: Imaging the Unsectioned Adult Zebra Finch Brain at Cellular Resolution

The inherent complexity of brain tissue, with brain cells intertwining locally and projecting to distant regions, has made three-dimensional visualization of intact brains a highly desirable but challenging task in neuroscience. The natural opaqueness of tissue has traditionally limited researchers...

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Bibliographic Details
Published in:Frontiers in neuroanatomy 2019-02, Vol.13, p.13-13
Main Authors: Rocha, Mariana Diales, Düring, Daniel Normen, Bethge, Philipp, Voigt, Fabian F, Hildebrand, Staffan, Helmchen, Fritjof, Pfeifer, Alexander, Hahnloser, Richard Hans Robert, Gahr, Manfred
Format: Article
Language:English
Subjects:
Anatomy
Animal models
Auditory discrimination learning
Brain architecture
Brain research
Cytomegalovirus
large volume imaging
light sheet microscopy
Magnetic resonance imaging
Microscopy
Nervous system
Neural networks
Neuroimaging
Neuroscience
Neurosciences
Sensorimotor system
Song
song system
tissue clearing
Vocalization behavior
zebra finch
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Summary:The inherent complexity of brain tissue, with brain cells intertwining locally and projecting to distant regions, has made three-dimensional visualization of intact brains a highly desirable but challenging task in neuroscience. The natural opaqueness of tissue has traditionally limited researchers to techniques short of single cell resolution such as computer tomography or magnetic resonance imaging. By contrast, techniques with single-cell resolution required mechanical slicing into thin sections, which entails tissue distortions that severely hinder accurate reconstruction of large volumes. Recent developments in tissue clearing and light sheet microscopy have made it possible to investigate large volumes at micrometer resolution. The value of tissue clearing has been shown in a variety of tissue types and animal models. However, its potential for examining the songbird brain remains unexplored. Songbirds are an established model system for the study of vocal learning and sensorimotor control. They share with humans the capacity to adapt vocalizations based on auditory input. Song learning and production are controlled in songbirds by the song system, which forms a network of interconnected discrete brain nuclei. Here, we use the CUBIC and iDISCO+ protocols for clearing adult songbird brain tissue. Combined with light sheet imaging, we show the potential of tissue clearing for the investigation of connectivity between song nuclei, as well as for neuroanatomy and brain vasculature studies.
ISSN:1662-5129
1662-5129
DOI:10.3389/fnana.2019.00013