Structural and molecular interrogation of intact biological systems.

Nature 2013 May 10;497(7449):332-7. Epub 2013 Apr 10.

Department of Bioengineering, Stanford University, Stanford, California 94305, USA.

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.

Download full-text PDF

Source
http://www.rna.uzh.ch/events/journalclub/20130423_JC_Gonde_n
Web Search
http://www.nature.com/doifinder/10.1038/nature12107
Publisher Site
http://dx.doi.org/10.1038/nature12107DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4092167PMC
May 2013
12 Reads

Publication Analysis

Top Keywords

clarity enables
8
human tissue
8
structural molecular
8
neurotransmitters clarity
4
acids neurotransmitters
4
complexes nucleic
4
protein complexes
4
enables intact-tissue
4
nucleic acids
4
situ hybridization
4
rounds staining
4
staining de-staining
4
de-staining non-sectioned
4
multiple rounds
4
immunohistochemistry multiple
4
structures protein
4
hybridization immunohistochemistry
4
intact-tissue situ
4
subcellular structures
4
macromolecule-permeable mouse
4

References

(Supplied by CrossRef)

W Denk et al.
Science 1990

P Carmeliet et al.
Nature 2005

F Helmchen et al.
Nature Methods 2005

W Denk et al.
PLoS Biol. 2004

KD Micheva et al.
Neuron 2007

J Livet et al.
Nature 2007

A Li et al.
Science 2010

EJ Botcherby et al.
Proc. Natl Acad. Sci. USA 2012

J DeFelipe et al.
Science 2010

CCH Petersen et al.
Neuron 2007

N Kasthuri et al.
Nature Methods 2007

Similar Publications