Structural and molecular interrogation of intact biological systems

Kwanghun Chung, Jenelle Wallace, Sung-Yon Kim, Sandhiya Kalyanasundaram, Aaron S. Andalman, Thomas J. Davidson, Julie J. Mirzabekov, Kelly A. Zalocusky, Joanna Mattis, Aleksandra K. Denisin, Sally Pak, Hannah Bernstein, Charu Ramakrishnan, Logan Grosenick, Viviana Gradinaru and Karl Deisseroth ()
Additional contact information
Kwanghun Chung: Stanford University
Jenelle Wallace: Stanford University
Sung-Yon Kim: Stanford University
Sandhiya Kalyanasundaram: CNC Program, Stanford University
Aaron S. Andalman: Stanford University
Thomas J. Davidson: Stanford University
Julie J. Mirzabekov: Stanford University
Kelly A. Zalocusky: Stanford University
Joanna Mattis: Stanford University
Aleksandra K. Denisin: Stanford University
Sally Pak: Stanford University
Hannah Bernstein: Stanford University
Charu Ramakrishnan: Stanford University
Logan Grosenick: Stanford University
Viviana Gradinaru: CNC Program, Stanford University
Karl Deisseroth: Stanford University

Nature, 2013, vol. 497, issue 7449, 332-337

Abstract: 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.

Date: 2013
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DOI: 10.1038/nature12107

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