AI Article Synopsis
- Understanding the structure-function relationships in biology requires detailed 3D maps of organs, which are not available for many species.
- The absence of whole-organ imaging methods has created a knowledge gap in anatomical and phenotypical understanding.
- This text introduces new techniques for tissue clearing, including PACT, RIMS, and PARS, enabling high-resolution imaging of intact organs and their cellular structures.
Article Abstract
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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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4153367 | PMC |
| http://dx.doi.org/10.1016/j.cell.2014.07.017 | DOI Listing |
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