Structural and molecular interrogation of intact biological systems

K Chung, J Wallace, SY Kim, S Kalyanasundaram… - Nature, 2013 - nature.com
K Chung, J Wallace, SY Kim, S Kalyanasundaram, AS Andalman, TJ Davidson
Nature, 2013nature.com
Obtaining high-resolution information from a complex system, while maintaining the global
perspective needed to understand system function, represents a key challenge in biology.
Here we address this challenge with a method (termed CLARITY) for the transformation of
intact tissue into a nanoporous hydrogel-hybridized form (crosslinked to a three-dimensional
network of hydrophilic polymers) that is fully assembled but optically transparent and
macromolecule-permeable. Using mouse brains, we show intact-tissue imaging of long …
Abstract
Obtaining high-resolution information from a complex system, while maintaining the global perspective needed to understand system function, represents a key challenge in biology. Here we address this challenge with a method (termed CLARITY) for the transformation of intact tissue into a nanoporous hydrogel-hybridized form (crosslinked to a three-dimensional network of hydrophilic polymers) that is fully assembled but optically transparent and macromolecule-permeable. Using mouse brains, we show intact-tissue imaging of long-range projections, local circuit wiring, cellular relationships, subcellular structures, protein complexes, nucleic acids and neurotransmitters. CLARITY also enables intact-tissue in situ hybridization, immunohistochemistry with multiple rounds of staining and de-staining in non-sectioned tissue, and antibody labelling throughout the intact adult mouse brain. Finally, we show that CLARITY enables fine structural analysis of clinical samples, including non-sectioned human tissue from a neuropsychiatric-disease setting, establishing a path for the transmutation of human tissue into a stable, intact and accessible form suitable for probing structural and molecular underpinnings of physiological function and disease.
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