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Clearing for Deep Tissue Imaging.

Authors
  • Muntifering, Michael1
  • Castranova, Daniel2
  • Gibson, Gregory A3
  • Meyer, Evan1
  • Kofron, Matthew1
  • Watson, Alan M3
  • 1 Division of Developmental Biology, Cincinnati Children's Research Foundation, Cincinnati, Ohio.
  • 2 Division of Developmental Biology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland.
  • 3 Center for Biologic Imaging, Department of Cell Biology, University of Pittsburgh, Pittsburgh, Pennsylvania.
Type
Published Article
Journal
Current protocols in cytometry
Publication Date
Oct 01, 2018
Volume
86
Issue
1
Identifiers
DOI: 10.1002/cpcy.38
PMID: 30005145
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Biologic tissues are generally opaque due to optical properties that result in scattering and absorption of light. Preparation of tissues for optical microscopy often involves sectioning to a thickness of 50-100 µm, the practical limits of light penetration and recovery. A researcher who wishes to image a whole tissue must acquire potentially hundreds of individual sections before rendering them into a three-dimensional volume. Clearing removes strongly light-scattering and light-absorbing components of a tissue and equalizes the refractive index of the imaging medium to that of the tissue. After clearing, the maximum depth of imaging is often defined by the microscope optics rather than the tissue. Such visibility enables the interrogation of whole tissues and even animals without the need to section. Researchers can study a biological process in the context of its three-dimensional environment, identify rare events in large volumes of tissues, and trace cells and cell-cell interactions over large distances. This article describes four popular clearing protocols that are relevant to a wide variety of scenarios across biologic disciplines: CUBIC, CLARITY, 3DISCO, and SeeDB. © 2018 by John Wiley & Sons, Inc. © 2018 John Wiley & Sons, Inc.

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