Simplified CLARITY for visualizing immunofluorescence labeling in the developing rat brain

Brain Structure & Function
H Zheng, Linda Rinaman

Abstract

CLARITY is an innovative technological advance in which intact biological tissue is transformed into a "nanoporous hydrogel-hybridized form" (Chung et al. 2013; Chung and Deisseroth 2013) with markedly improved chemical and optical accessibility, permitting fluorescent visualization and extraction of high-resolution structural data from mm-thick blocks of tissue. CLARITY affords an excellent 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 polymerization 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 equipp...Continue Reading

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Citations

Jul 22, 2017·Journal of Chemical Neuroanatomy·Kristian H Reveles Jensen, Rune W Berg
May 25, 2018·Annual Review of Biophysics·Viviana GradinaruKarl Deisseroth
Dec 27, 2017·Journal of Biophotonics·Paweł MatrybaMarzena Stefaniuk
Feb 13, 2018·Cellular & Molecular Immunology·David Twapokera MzinzaReinhold Förster
Oct 30, 2016·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Huixia RenHuanxing Su
Sep 7, 2018·Experimental and Therapeutic Medicine·Hao DuQiyu Li
Jun 4, 2020·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Daniel Smith ParedesBhart-Anjan S Bhullar
Dec 29, 2020·Journal of Neuroscience Methods·Guadalupe Martínez-LorenzanaMiguel Condés-Lara
Dec 17, 2020·ACS Chemical Neuroscience·Jiajia ZhaoHaitao Sun

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