Looking through Brains with Fast Passive CLARITY: Zebrafish, Rodents, Non-human Primates and Humans

Recently developed CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ-hybridization-compatible Tis-sue-hYdrogel) technique renders the tissue transparent by removing lipids in the tissue, while preserving and stabilizing the cellular and subcellular structures....

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Bibliographic Details
Published inBio-protocol Vol. 9; no. 15; p. e3321
Main Authors Mortazavi, Farzad, Stankiewicz, Alexander J, Zhdanova, Irina V
Format Journal Article
LanguageEnglish
Published United States Bio-Protocol 05.08.2019
Bio-protocol LLC
Subjects
Biology
Methods
Human
Immunohistochemistry
Brain
Rhesus monkey
Rat
Adult neurogenesis
Imaging
Whole-tissue staining
Non-human primate
CLARITY
Zebrafish
Mice
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Summary:Recently developed CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging/Immunostaining/In situ-hybridization-compatible Tis-sue-hYdrogel) technique renders the tissue transparent by removing lipids in the tissue, while preserving and stabilizing the cellular and subcellular structures. This provides effective penetration of diverse labeling probes, from primary and secondary antibodies to complementary DNA and RNA strands. Followed by high-resolution 3D imaging of neuronal cells and their projections in thick sections, tissue blocks, whole brains, or whole animals, CLARITY allows for superior quantitative analysis of neuronal tissue. Here, we provide our detailed protocol for PACT (Passive Clarity Technique) in brain tissue of diverse species, including human, non-human primate, rodents, and zebrafish. We describe the six principal steps: (1) Tissue fixation and preparation, (2) Passive lipid removal, (3) Immuno-labeling, (4) Optical clearing, (5) Imaging, (6) 3D visualization and quantification.
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ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.3321