AI Article Synopsis
- A new set of tissue fixation and clearing protocols allows for the transformation of organs and whole organisms into clear samples in 1-2 weeks, preserving cellular structure and fluorescence.
- The techniques PACT and PARS utilize tissue-hydrogel hybrids to improve the clarity and integrity of samples by facilitating lipid removal, making it easier for imaging agents to penetrate the tissue.
- These methods are versatile, applicable to challenging tissues like bone, and enhance imaging capabilities, allowing for detailed visualization of cellular components and networks.
Article Abstract
To facilitate fine-scale phenotyping of whole specimens, we describe here a set of tissue fixation-embedding, detergent-clearing and staining protocols that can be used to transform excised organs and whole organisms into optically transparent samples within 1-2 weeks without compromising their cellular architecture or endogenous fluorescence. PACT (passive CLARITY technique) and PARS (perfusion-assisted agent release in situ) use tissue-hydrogel hybrids to stabilize tissue biomolecules during selective lipid extraction, resulting in enhanced clearing efficiency and sample integrity. Furthermore, the macromolecule permeability of PACT- and PARS-processed tissue hybrids supports the diffusion of immunolabels throughout intact tissue, whereas RIMS (refractive index matching solution) grants high-resolution imaging at depth by further reducing light scattering in cleared and uncleared samples alike. These methods are adaptable to difficult-to-image tissues, such as bone (PACT-deCAL), and to magnified single-cell visualization (ePACT). Together, these protocols and solutions enable phenotyping of subcellular components and tracing cellular connectivity in intact biological networks.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4917295 | PMC |
| http://dx.doi.org/10.1038/nprot.2015.122 | DOI Listing |
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