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Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping

Nature protocols, 2015-11, Vol.10 (11), p.1860-1896 [Peer Reviewed Journal]

COPYRIGHT 2015 Nature Publishing Group ;COPYRIGHT 2015 Nature Publishing Group ;Copyright Nature Publishing Group Nov 2015 ;Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. 2015. ;ISSN: 1754-2189 ;EISSN: 1750-2799 ;DOI: 10.1038/nprot.2015.122 ;PMID: 26492141

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Title:
Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping
Author: Treweek, Jennifer B ; Chan, Ken Y ; Flytzanis, Nicholas C ; Yang, Bin ; Deverman, Benjamin E ; Greenbaum, Alon ; Lignell, Antti ; Xiao, Cheng ; Cai, Long ; Ladinsky, Mark S ; Bjorkman, Pamela J ; Fowlkes, Charless C ; Gradinaru, Viviana
Subjects: Analysis ; Animals ; Biological research ; Biological specimens ; Biology, Experimental ; Biomolecules ; Caenorhabditis elegans ; Cellular communication ; Cellular signal transduction ; Circuits ; Clearing ; Detergents - isolation & purification ; Embedding ; Extraction (Chemistry) ; Fluorescence ; Fluorescence microscopy ; Genetic aspects ; High resolution ; Histocytochemistry - methods ; Hybrids ; Hydrogels ; Image resolution ; Light scattering ; Lipids ; Lipids - isolation & purification ; Methods ; Mice ; Optical Imaging - methods ; Organs ; Pathology - methods ; Perfusion ; Permeability ; Phenotype ; Phenotyping ; Physiological aspects ; Rats ; Refractivity ; Specimen Handling - methods ; Staining and Labeling - methods ; Time Factors ; Tissue Embedding - methods ; Tissue Fixation - methods ; Tissues
Is Part Of: Nature protocols, 2015-11, Vol.10 (11), p.1860-1896
Description: 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.
Publisher: England: Nature Publishing Group
Language: English
Identifier: ISSN: 1754-2189
EISSN: 1750-2799
DOI: 10.1038/nprot.2015.122
PMID: 26492141
Source: MEDLINE
ProQuest Central

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Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping

COPYRIGHT 2015 Nature Publishing Group ;COPYRIGHT 2015 Nature Publishing Group ;Copyright Nature Publishing Group Nov 2015 ;Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. 2015. ;ISSN: 1754-2189 ;EISSN: 1750-2799 ;DOI: 10.1038/nprot.2015.122 ;PMID: 26492141

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