🏆 Foundational Paper

Whole-Brain Profiling of Cells and Circuits in Mammals by Tissue Clearing and Light-Sheet Microscopy.

Ueda Hiroki R, Dodt Hans-Ulrich, Osten Pavel, Economo Michael N, Chandrashekar Jayaram, Keller Philipp J

📰 Neuron 📅 2020 📊 175 citations
DOI PubMed

Abstract

Tissue clearing and light-sheet microscopy have a 100-year-plus history, yet these fields have been combined only recently to facilitate novel experiments and measurements in neuroscience. Since tissue-clearing methods were first combined with modernized light-sheet microscopy a decade ago, the performance of both technologies has rapidly improved, broadening their applications. Here, we review the state of the art of tissue-clearing methods and light-sheet microscopy and discuss applications of these techniques in profiling cells and circuits in mice. We examine outstanding challenges and future opportunities for expanding these techniques to achieve brain-wide profiling of cells and circuits in primates and humans. Such integration will help provide a systems-level understanding of the physiology and pathology of our central nervous system.

🔬 Techniques

Confocal Laser Scanning Microscopy Expansion Microscopy Fluorescence Microscopy Immunofluorescence Microscopy Light Sheet Fluorescence Microscopy Two-Photon Excitation Microscopy

🔭 Microscopes

LaVision BioTec UltraMicroscope Light Sheet UltraMicroscope

🧬 Organisms

Caenorhabditis elegans Xenopus laevis

💻 Software

OMERO

✨ Fluorophores

GFP mCherry

🧪 Sample Preparation

Adeno-Associated Virus Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging-compatible Tissue-hydrogel Clear Unobstructed Brain Imaging Cocktails and Computational analysis Expansion Microscopy Fixation Fluorescence In Situ Hybridization Immunofluorescence Immunolabeling-enabled Three-Dimensional Imaging of Solvent-Cleared Organs Immunostaining In Situ Hybridization Live Imaging Permeabilization Polyethylene Glycol-Associated Solvent System Three-Dimensional Imaging of Solvent-Cleared Organs Tissue Clearing Transduction Ultimate Three-Dimensional Imaging of Solvent-Cleared Organs Whole Mount

🔬 Cell Lines

Primary Microglia

🏭 Microscope Brands

LaVision BioTec

📷 Detectors

CCD sCMOS

💻 Software Details

Image Analysis:
OMERO

🏛️ Research Organizations (ROR)

Affiliated research institutions:

🏛 The University of Tokyo 🏛 RIKEN 🏛 Medical University of Vienna 🏛 University of Vienna 🏛 Modelica Association 🏛 Boston University 🏛 Janelia Research Campus 🏛 Howard Hughes Medical Institute 🏛 TU Wien 🏛 BioElectronics (United States) 🏛 Cold Spring Harbor Laboratory 🏛 United States Department of Veterans Affairs

📊 Figures

Figure 1.

Figure 1.

Timeline of Advances in Tissue Clearing Methods.

Figure 2.

Figure 2.

Physical and Chemical Principles of Tissue Clearing.

(A) Classical Physical principles of tissue clearing proposed in 1911 by Spalteholz. Light scattering can be minimized by homogenization of RIs of materials whereas light absorption can be minimized b...

Figure 3.

Figure 3.

Timeline of Advances in Light-Sheet Microscopy.

Figure 4.

Figure 4.

Whole-Brain Profiling of Cells by Light-Sheet Fluorescent Microscopy.

(A) Volume-rendered and single plane images of a brain transduced with AAV-PHP.eB:NSE-H2B-mCherry (mCherry, green) counterstained by RD2 (red) which is cleared by CUBIC-L/R+. Overlapped signals are sh...

Figure 5.

Figure 5.

Whole-Brain Profiling of Circuits.

Single-neuron reconstruction reveals structured connectivity patterns. (A) Left, Anterograde tracing of a population of neurons in a source brain region can reveal regions of the brain to which the so...

Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.

🏛️ Imaging Facility

🏛️ University of Tokyo

💬 Discussion

0 comments

No comments yet. Be the first to start a discussion!

Leave a Comment

MicroHub Assistant