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Cited by 421Single-cell and spatial transcriptomicsAdvanced Fluorescence Microscopy TechniquesClick Chemistry and Applications

Protection of tissue physicochemical properties using polyfunctional crosslinkers

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Park, Young-Gyun, Sohn, Chang Ho, Chen, Ritchie, McCue, Margaret, Yun, Dae Hee, Drummond, Gabrielle T, Ku, Taeyun, Evans, Nicholas B +15 more·2019·Nature Biotechnology
DOI 10.1038/nbt.4281PMC PMC6579717
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Abstract

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.

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  • ROI delineation involved cycles of interactions between automated protocols and appraisal by skilled neuroanatomists following the strategy applied to human high throughput neuroimaging.

    “In brief, this has involved cycles of interactions between automated protocols described above and an appraisal by skilled neuroanatomists following the strategy applied to human high throughput neuroimaging95.”

    From Magnetic resonance microscopy maps widespread effects of Alzheimer’s disease on brain structures and behavior in mice (2026) · cites this paper as [95]

    The Materials and Methods sections describe automatic atlas alignment and human-reviewed, semi-automatic axon tracing, but not the claimed strategy for ROI delineation in human high-throughput neuroimaging. Searches for skilled neuroanatomist appraisal and iterative automated ROI delineation found no passage addressing that strategy. (LLM, conf=0.73)

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Authors
Park, Y.-G., Sohn, C. H., Chen, R., McCue, M., Yun, D. H., Drummond, G. T., Ku, T., Evans, N. B., Oak, H. C., Trieu, W., Choi, H., Jin, X., Lilascharoen, V., Wang, J., Truttmann, M. C., Qi, H. W., Ploegh, H. L., Golub, T. R., Chen, S.-C., Frosch, M. P., Kulik, H. J., Lim, B. K., & Chung, K.
Affiliations
  • Massachusetts Institute of Technology (US)
  • Broad Institute (US)
  • University of California San Diego (US)
  • Chinese University of Hong Kong (HK)
  • Boston Children's Hospital (US)
  • Harvard University (US)
Corresponding author
Chung, Kwanghun
ORCID
Venue
Nature Biotechnology
Publisher
Springer Science and Business Media LLC
Published
2018-12-17
Volume
37
Issue
1
Pages
73-83
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
DOI
10.1038/nbt.4281
PMC
PMC6579717
OpenAlex
W2904533595
Cited by
421
References
3
Citations by year
2026: 54 · 2025: 72 · 2024: 77 · 2023: 57 · 2022: 48 · 2021: 62 · 2020: 35 · 2019: 15 · 2018: 1
FWCI
9.87
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc)
  • eScholarship (California Digital Library) (submittedVersion)
  • PMC
Funders
  • National Science Foundation (1429830, ACI-1 548 562, 1548562)
  • U.S. Department of Energy (1548562, ACI-1548562)
  • Burroughs Wellcome Fund
  • McKnight Foundation
  • National Alliance for Research on Schizophrenia and Depression
  • Searle Scholars Program
  • National Institutes of Health (1548562, ACI-1548562, AG005134)
  • Simons Center for the Social Brain, Massachusetts Institute of Technology
  • National Institute on Aging (AG005134, P50AG005134)
  • National Institute of Mental Health (R01MH108594, U01MH114829, R01MH107742)
  • National Institute of Environmental Health Sciences (DP2ES027992)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
Keywords
tissue stabilization, biomolecule preservation, tissue architecture preservation, synaptic structure, human brain cells, tissue transformation

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