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- W3151861713 abstract "CLARITY is an innovative technological ad- vance in which intact biological tissue is transformed into a ''nanoporous hydrogel-hybridized form'' (Chung et al. 2013; Chung and Deisseroth 2013) with markedly improved che- mical and optical accessibility, permitting fluorescent visu- alization and extraction of high-resolution structural data from mm-thick blocks of tissue. CLARITY affords an ex- cellent but as yet unexploited opportunity to visualize the growth and maturation of phenotypically identified neurons and axonal processes in the developing brain. This brief report describes a moderately revised, simplified, and less expensive CLARITY protocol that effectively reveals the structure of chemically identified neurons in whole neonatal/ juvenile rat brains and tissue slabs. Rats (postnatal day (P)0-24) were transcardially perfused with one of two fixative/hydrogel solutions, followed by hydrogel polymer- ization to generate brain hybrids. Whole brain hybrids or 2.0-mm-thick coronal slabs were passively cleared of lipid and then processed for dual immunofluorescence labeling, including labeling using tyramide signal amplification. After refractive index matching using 2,20-Thiodiethanol (60 % solution), a Leica confocal microscope equipped with a CLARITY objective was used to view the hypothalamus in whole brain hybrids or slabs. Collected image stacks re- vealed the distribution and three-dimensional structure of hypothalamic pro-oxyphysin (oxytocin)-, neuropeptide Y-, glucagon-like peptide-1-, and tyrosine hydroxylase- immunopositive neurons and processes within large tissue volumes. Outstanding structural preservation and immuno- labeling quality demonstrates the efficacy of this approach for interrogating chemically defined neural circuits as they develop in postnatal rodent brain." @default.
- W3151861713 created "2021-04-13" @default.
- W3151861713 creator A5017713926 @default.
- W3151861713 date "2016-01-01" @default.
- W3151861713 modified "2023-09-27" @default.
- W3151861713 title "Simplified CLARITY for visualizing immunofluorescence labeling in the developing rat brain" @default.
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