Journal
CELL
Volume 158, Issue 4, Pages 945-958Publisher
CELL PRESS
DOI: 10.1016/j.cell.2014.07.017
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Funding
- NIH/NINDS New Innovator [NIH IDP20D017782-01]
- NIH [1R01AG047664-01, R01HD075605, 1R01NS085910-01, 1R21MH103824-01]
- Startup funds from the President and Provost of California Institute of Technology
- Biology and Biological Engineering Division of California Institute of Technology
- Beckman Institute of Caltech
- Pew Charitable Trust
- Sidney Kimmel Foundation
- Human Frontiers in Science Program
- Mallinckrodt Foundation
- Gordon and Betty Moore Foundation
- Michael J. Fox Foundation
- Caltech-GIST
- Caltech Biology Division Training grant [NIH/NRSA 5T32GM07616]
- NIH/NIAMS [5T32AR058921]
- Colvin Postdoctoral Fellowship
- CALTECH Division of BBE
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Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Herein, we present techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity. We describe PACT (passive clarity technique), a protocol for passive tissue clearing and immunostaining of intact organs; RIMS (refractive index matching solution), a mounting media for imaging thick tissue; and PARS (perfusion-assisted agent release in situ), a method for whole-body clearing and immunolabeling. We show that in rodents PACT, RIMS, and PARS are compatible with endogenous-fluorescence, immunohistochemistry, RNA single-molecule FISH, long-term storage, and microscopy with cellular and subcellular resolution. These methods are applicable for high-resolution, high-content mapping and phenotyping of normal and pathological elements within intact organs and bodies.
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