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

Taeyun Ku, Justin Swaney, Jeong Yoon Park, Alexandre Albanese, Evan Murray, Jae Hun Cho, Young Gyun Park, Vamsi Mangena, Jiapei Chen, Kwanghun Chung

Research output: Contribution to journalArticlepeer-review

134 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)973-981
Number of pages9
JournalNature Biotechnology
Volume34
Issue number9
DOIs
Publication statusPublished - 2016 Sep 1

Bibliographical note

Funding Information:
The authors thank the entire Chung laboratory for support and helpful discussions. K.C. was supported by Burroughs Wellcome Fund Career Awards at the Scientific Interface, the Searle Scholars Program, Packard award in Science and Engineering, JPB Foundation (PIIF and PNDRF) and NIH (1-U01-NS090473-01).

Publisher Copyright:
© 2016 Nature America, Inc.

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

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