Chemical Landscape for Tissue Clearing Based on Hydrophilic Reagents

Cell Reports, Journal Year: 2018, Volume and Issue: 24(8), P. 2196 - 2210.e9

Published: Aug. 1, 2018

We describe a strategy for developing hydrophilic chemical cocktails tissue delipidation, decoloring, refractive index (RI) matching, and decalcification, based on comprehensive profiling. More than 1,600 chemicals were screened by high-throughput evaluation system each process. The profiling revealed important factors: salt-free amine with high octanol/water partition-coefficient (logP) N-alkylimidazole aromatic amide RI protonation of phosphate ion decalcification. strategic integration optimal provided series CUBIC (clear, unobstructed brain/body imaging computational analysis) protocols, which efficiently clear mouse organs, body including bone, even large primate human tissues. updated protocols are scalable reproducible, they enable three-dimensional the mammalian This represents future paradigm rational design clearing that can be used

Language: Английский

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The emerging landscape of spatial profiling technologies

Nature Reviews Genetics, Journal Year: 2022, Volume and Issue: 23(12), P. 741 - 759

Published: July 20, 2022

Language: Английский

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Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues

Nature Communications, Journal Year: 2020, Volume and Issue: 11(1)

Published: April 27, 2020

Abstract Whole-organ/body three-dimensional (3D) staining and imaging have been enduring challenges in histology. By dissecting the complex physicochemical environment of system, we developed a highly optimized 3D pipeline based on CUBIC. Based our precise characterization biological tissues as an electrolyte gel, experimentally evaluated broad conditions by using artificial tissue-mimicking material. The combination allows bottom-up design superior protocol that can uniformly label whole adult mouse brains, marmoset brain hemisphere, ~1 cm 3 tissue block postmortem human cerebellum, entire infant body with dozens antibodies cell-impermeant nuclear stains. whole-organ images collected light-sheet microscopy are used for computational analyses comparison analysis between species. This pipeline, named CUBIC-HistoVIsion, thus offers advanced opportunities organ- organism-scale histological multicellular systems.

Language: Английский

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Molecular imaging of biological systems with a clickable dye in the broad 800- to 1,700-nm near-infrared window

Proceedings of the National Academy of Sciences, Journal Year: 2017, Volume and Issue: 114(5), P. 962 - 967

Published: Jan. 17, 2017

Significance Fluorescence-based optical imaging is an important tool allowing researchers and clinicians to molecularly probe wide-ranging biological structures processes. To break through the traditional molecular window spanning from visible near-infrared (NIR)-I (400–900 nm) region for multiplicity, newly designed ultrapurified fluorescent probe-antibody conjugates with fluorescence emissions in NIR-II (1,000–1,700 have been developed. These probes can reduce background autofluorescence deep-tissue a 3D mode. open up more deeper nonoverlapping channels complex systems.

Language: Английский

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Chemical Principles in Tissue Clearing and Staining Protocols for Whole-Body Cell Profiling

Annual Review of Cell and Developmental Biology, Journal Year: 2016, Volume and Issue: 32(1), P. 713 - 741

Published: June 14, 2016

Mammalian bodies have more than a billion cells per cubic centimeter, which makes whole-body cell (WBC) profiling of an organism one the ultimate challenges in biology and medicine. Recent advances tissue-clearing technology enabled rapid comprehensive cellular analyses whole organs body by combination state-of-the-art technologies optical imaging image informatics. In this review, we focus mainly on chemical principles currently available techniques for tissue clearing staining to facilitate our understanding their underlying mechanisms. Tissue is usually conducted following steps: (a) fixation, (b) permeabilization, (c) decolorizing, (d) refractive index (RI) matching. To phenotype individual after clearing, it important visualize genetically encoded fluorescent reporters and/or stain tissues with dyes, labeled antibodies, or nucleic acid probes. Although some technical remain, WBC will enable various applications, such as identifying circuits across multiple measuring dynamics stochastic proliferative processes, example, autoimmune malignant neoplastic diseases.

Language: Английский

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Spatial mapping of protein composition and tissue organization: a primer for multiplexed antibody-based imaging

Nature Methods, Journal Year: 2021, Volume and Issue: 19(3), P. 284 - 295

Published: Nov. 22, 2021

Language: Английский

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Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (C _e 3D)

Proceedings of the National Academy of Sciences, Journal Year: 2017, Volume and Issue: 114(35)

Published: Aug. 14, 2017

Significance Major biological processes rely on the precise positioning of diverse cell types in specific anatomical locations. Existing techniques for studying cellular spatial tissues, especially with robust identification densely packed cells, have substantial time, cost, resolution, and multiplexing limitations. Here, we describe an easy-to-use inexpensive tissue clearing technique attaining high-quality images cells molecules interest volumes, enabling simultaneous quantitative analysis 3D organ structure fine-grained composition. This technology will enhance our capacity acquiring a understanding relationships between their microenvironments context broader organization is directly applicable to disciplines as well diagnostic medicine.

Language: Английский

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Cellular and Molecular Probing of Intact Human Organs

Cell, Journal Year: 2020, Volume and Issue: 180(4), P. 796 - 812.e19

Published: Feb. 1, 2020

Language: Английский

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Three-Dimensional Study of Alzheimer’s Disease Hallmarks Using the iDISCO Clearing Method

Cell Reports, Journal Year: 2016, Volume and Issue: 16(4), P. 1138 - 1152

Published: July 1, 2016

Amyloidosis is a major problem in over one hundred diseases, including Alzheimer's disease (AD). Using the iDISCO visualization method involving targeted molecular labeling, tissue clearing, and light-sheet microscopy, we studied plaque formation intact AD mouse brain at up to 27 months of age. We visualized amyloid plaques 3D together with tau, microglia, vasculature. Volume imaging coupled automated detection mapping enables precise fast quantification within entire brain. The present methodology also applicable analysis frozen human samples without specialized preservation. Remarkably, tissues showed greater complexity surprisingly large three-dimensional patterns, or TAPs. ability visualize 3D, especially context their micro-environment, discovery TAPs may have important scientific medical implications.

Language: Английский

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

Neuron, Journal Year: 2020, Volume and Issue: 106(3), P. 369 - 387

Published: May 1, 2020

Language: Английский

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