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Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues

The biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The me... Full description

Journal Title: Nature biotechnology 2016-09, Vol.34 (9), p.973-981
Main Author: Ku, Taeyun
Other Authors: Swaney, Justin , Park, Jeong-Yoon , Albanese, Alexandre , Murray, Evan , Cho, Jae Hun , Park, Young-Gyun , Mangena, Vamsi , Chen, Jiapei , Chung, Kwanghun
Format: Electronic Article Electronic Article
Language: English
Subjects:
Publisher: United States: Nature Publishing Group
ID: ISSN: 1087-0156
Link: https://www.ncbi.nlm.nih.gov/pubmed/27454740
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recordid: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5070610
title: Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues
format: Article
creator:
  • Ku, Taeyun
  • Swaney, Justin
  • Park, Jeong-Yoon
  • Albanese, Alexandre
  • Murray, Evan
  • Cho, Jae Hun
  • Park, Young-Gyun
  • Mangena, Vamsi
  • Chen, Jiapei
  • Chung, Kwanghun
subjects:
  • Animals
  • Article
  • Biology
  • Brain - metabolism
  • Brain - ultrastructure
  • Female
  • Gene Expression Profiling - methods
  • Gene loci
  • Image Enhancement - methods
  • Image Interpretation, Computer-Assisted - methods
  • Imaging, Three-Dimensional - methods
  • Immunoassay - methods
  • Male
  • Medical imaging
  • Methods
  • Mice
  • Molecular Imaging - methods
  • Nerve Tissue Proteins - metabolism
  • Observations
  • Protein-protein interactions
  • Proteins
  • Proteome - metabolism
  • Proteome - ultrastructure
  • Proteomics
  • Synapses - metabolism
  • Synapses - ultrastructure
  • Tissue Distribution
  • Tissues
ispartof: Nature biotechnology, 2016-09, Vol.34 (9), p.973-981
description: The biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.
language: eng
source:
identifier: ISSN: 1087-0156
fulltext: no_fulltext
issn:
  • 1087-0156
  • 1546-1696
url: Link


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descriptionThe biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.
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subjectAnimals ; Article ; Biology ; Brain - metabolism ; Brain - ultrastructure ; Female ; Gene Expression Profiling - methods ; Gene loci ; Image Enhancement - methods ; Image Interpretation, Computer-Assisted - methods ; Imaging, Three-Dimensional - methods ; Immunoassay - methods ; Male ; Medical imaging ; Methods ; Mice ; Molecular Imaging - methods ; Nerve Tissue Proteins - metabolism ; Observations ; Protein-protein interactions ; Proteins ; Proteome - metabolism ; Proteome - ultrastructure ; Proteomics ; Synapses - metabolism ; Synapses - ultrastructure ; Tissue Distribution ; Tissues
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notesThese authors contributed equally to this work.
abstractThe biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.
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