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

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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.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2016-09
Language
English
Article Type
Article
Citation

NATURE BIOTECHNOLOGY, v.34, no.9, pp.973 - +

ISSN
1087-0156
DOI
10.1038/nbt.3641
URI
http://hdl.handle.net/10203/264050
Appears in Collection
MSE-Journal Papers(저널논문)
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