Protection of tissue physicochemical properties using polyfunctional crosslinkers

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Understanding complex biological systems requires the system-wide characterization of both molecular and cellular features. Existing methods for spatial mapping of biomolecules in intact tissues suffer from information loss caused by degradation and tissue damage. We report a tissue transformation strategy named stabilization under harsh conditions via intramolecular epoxide linkages to prevent degradation (SHIELD), which uses a flexible polyepoxide to form controlled intra- and intermolecular cross-link with biomolecules. SHIELD preserves protein fluorescence and antigenicity, transcripts and tissue architecture under a wide range of harsh conditions. We applied SHIELD to interrogate system-level wiring, synaptic architecture, and molecular features of virally labeled neurons and their targets in mouse at single-cell resolution. We also demonstrated rapid three-dimensional phenotyping of core needle biopsies and human brain cells. SHIELD enables rapid, multiscale, integrated molecular phenotyping of both animal and clinical tissues.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2019-01
Language
English
Article Type
Article
Citation

NATURE BIOTECHNOLOGY, v.37, no.1, pp.73 - 83

ISSN
1087-0156
DOI
10.1038/nbt.4281
URI
http://hdl.handle.net/10203/269104
Appears in Collection
MSE-Journal Papers(저널논문)
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