A versatile depigmentation, clearing, and labeling method for exploring nervous system diversity.

Science Advances
Marko PendeHans-Ulrich Dodt

Abstract

Tissue clearing combined with deep imaging has emerged as a powerful alternative to classical histological techniques. Whereas current techniques have been optimized for imaging selected nonpigmented organs such as the mammalian brain, natural pigmentation remains challenging for most other biological specimens of larger volume. We have developed a fast DEpigmEntation-Plus-Clearing method (DEEP-Clear) that is easily incorporated in existing workflows and combines whole system labeling with a spectrum of detection techniques, ranging from immunohistochemistry to RNA in situ hybridization, labeling of proliferative cells (EdU labeling) and visualization of transgenic markers. With light-sheet imaging of whole animals and detailed confocal studies on pigmented organs, we provide unprecedented insight into eyes, whole nervous systems, and subcellular structures in animal models ranging from worms and squids to axolotls and zebrafish. DEEP-Clear thus paves the way for the exploration of species-rich clades and developmental stages that are largely inaccessible by regular imaging approaches.

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Citations

Oct 18, 2020·Proceedings of the National Academy of Sciences of the United States of America·Eric J KochMargaret McFall-Ngai
Oct 22, 2020·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Cristina Subiran AdradosMaximina Hee Yun
Dec 19, 2020·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Wouter Masselink, Elly M Tanaka
Jan 8, 2021·PLoS Biology·Bruno M FontinhaKristin Tessmar-Raible
Oct 30, 2020·Frontiers in Immunology·Elisa Eme-Scolan, Samantha J Dando
Feb 21, 2021·Neuron·Nicholas Jourjine, Hopi E Hoekstra

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Methods Mentioned

BETA
confocal microscopy
transgenic
light-sheet
confocal
electron microscopy
light microscopy
imaging technique

Software Mentioned

DEEP
FlyClear
Clear

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