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Nat Biotechnol. 2016 Sep;34(9):973-81. doi: 10.1038/nbt.3641. Epub 2016 Jul 25.

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

Author information

1
Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.
2
Picower Institute for Learning and Memory, MIT, Cambridge, Massachusetts, USA.
3
Department of Chemical Engineering, MIT, Cambridge, Massachusetts, USA.
4
Department of Neurosurgery, Gangnam Severance Hospital, Spine and Spinal Cord Institute, Yonsei University College of Medicine, Seoul, Republic of Korea.
5
Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA.
6
Harvard-MIT Division of Health Sciences and Technology, MIT, Cambridge, Massachusetts, USA.
7
Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA.
8
Broad Institute of Harvard University and MIT, Cambridge, Massachusetts, USA.

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.

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PMID:
27454740
PMCID:
PMC5070610
DOI:
10.1038/nbt.3641
[Indexed for MEDLINE]
Free PMC Article

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