Clearing for Deep Tissue Imaging

Current Protocols in Cytometry
Michael MuntiferingAlan M Watson

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.

References

Oct 13, 2012·Nature Protocols·Ali ErtürkHans-Ulrich Dodt
Apr 12, 2013·Nature·Kwanghun ChungKarl Deisseroth
Jun 1, 2013·Nature Methods·Kwanghun Chung, Karl Deisseroth
Mar 12, 2015·Frontiers in Neuroanatomy·Bing HouTianzi Jiang
Jul 18, 2015·Cell·Douglas S Richardson, Jeff W Lichtman
Oct 9, 2015·Nature Protocols·Etsuo A SusakiHiroki R Ueda
May 5, 2016·Progress in Histochemistry and Cytochemistry·Adriano AzaripourCornelis J F Van Noorden
Aug 23, 2016·Nature Methods·Chenchen PanAli Ertürk

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Citations

Nov 1, 2018·Current Protocols in Cytometry·Piyusha S GadeChih-Yuan Chuang
Aug 6, 2020·Journal of the Royal Society, Interface·Jeremy D Eekhoff, Spencer P Lake
May 28, 2021·Frontiers in Endocrinology·Michael P Dybala, Manami Hara
Jun 25, 2021·Advanced Science·Joanne X ShenVolker M Lauschke
Jul 3, 2021·Vaccines·Matthieu Van TilbeurghCaroline Manet

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