Structural and molecular interrogation of intact biological systems

Nature. 2013 May 16;497(7449):332-7. doi: 10.1038/nature12107. Epub 2013 Apr 10.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Brain / anatomy & histology*
  • Cross-Linking Reagents / chemistry
  • Formaldehyde / chemistry
  • Humans
  • Hydrogel, Polyethylene Glycol Dimethacrylate / chemistry
  • Imaging, Three-Dimensional / methods*
  • In Situ Hybridization / methods
  • Lipids / isolation & purification
  • Mice
  • Molecular Imaging / methods*
  • Permeability
  • Phenotype
  • Scattering, Radiation

Substances

  • Cross-Linking Reagents
  • Lipids
  • Formaldehyde
  • Hydrogel, Polyethylene Glycol Dimethacrylate