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Simultaneous protection of tissue physicochemical properties using polyfunctional crosslinkers

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 s... Full description

Journal Title: Nature biotechnology 2018-12-01
Main Author: Park, Young-Gyun
Other Authors: Sohn, Chang Ho , Chen, Ritchie , Mccue, Margaret , Yun, Dae Hee , Drummond, Gabrielle T. , Ku, Taeyun , Evans, Nicholas B. , Oak, Hayeon Caitlyn , Trieu, Wendy , Choi, Heejin , Jin, Xin , Lilascharoen, Varoth , Wang, Ji , Truttmann, Matthias C. , Qi, Helena W. , Ploegh, Hidde L. , Golub, Todd R. , Chen, Shih-Chi , Frosch, Matthew P. , Kulik, Heather J. , Lim, Byung Kook , Chung, Kwanghun
Format: Electronic Article Electronic Article
Language: English
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ID: ISSN: 1087-0156
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title: Simultaneous protection of tissue physicochemical properties using polyfunctional crosslinkers
format: Article
creator:
  • Park, Young-Gyun
  • Sohn, Chang Ho
  • Chen, Ritchie
  • Mccue, Margaret
  • Yun, Dae Hee
  • Drummond, Gabrielle T.
  • Ku, Taeyun
  • Evans, Nicholas B.
  • Oak, Hayeon Caitlyn
  • Trieu, Wendy
  • Choi, Heejin
  • Jin, Xin
  • Lilascharoen, Varoth
  • Wang, Ji
  • Truttmann, Matthias C.
  • Qi, Helena W.
  • Ploegh, Hidde L.
  • Golub, Todd R.
  • Chen, Shih-Chi
  • Frosch, Matthew P.
  • Kulik, Heather J.
  • Lim, Byung Kook
  • Chung, Kwanghun
subjects:
  • Article
ispartof: Nature biotechnology, 2018-12-01
description: 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 crosslink with biomolecules. SHIELD preserved 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 (3D) phenotyping of core needle biopsies and human brain cells. SHIELD enables rapid, multiscale, integrated molecular phenotyping of both animal and clinical tissues.
language: eng
source:
identifier: ISSN: 1087-0156
fulltext: no_fulltext
issn:
  • 1087-0156
  • 1546-1696
url: Link


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titleSimultaneous protection of tissue physicochemical properties using polyfunctional crosslinkers
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descriptionUnderstanding 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 crosslink with biomolecules. SHIELD preserved 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 (3D) 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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0AUTHOR CONTRIBUTIONS
1Y.-G.P., C.H.S., R.C., D.H.Y., and K.C. designed the study and wrote the paper with input from other authors. Y.-G.P. led development of SHIELD-SWITCH and SHIELD-MAP methods. C.H.S. and R.C. designed and performed the FISH experiments and protein analysis. R.C. and M.M. performed the fluorescence protein experiments. Y.-G.P., C.H.S., and M.M. performed the immunoreactivity experiments. Y.-G.P. and C.H.S. characterized physical properties of SHIELD tissue and performed axon tracing. D.H.Y. carried out active clearing and staining. D.H.Y. performed 3D phenotyping of the biopsy samples. G.T.D. and C.H.S. developed the SHIELD protocol for postmortem human brain tissues and performed immunostaining and imaging of the human tissues. T.K. performed the tissue expansion analysis. N.B.E. conducted light-sheet microscope imaging. H.C.O. and W.T. helped sample preparation. H.C. built temporally focused line-scanning microscope. M.C.T. and H.L.P. provided purified GFP. X.J. and T.R.G. provided tumor tissues. S.-C.C. and J.W. developed the oscillating blade microtome. M.P.F provided human brain tissues and validated the human brain data. H.J.K and H.W.Q. performed molecular simulation. V.L. and B.K.L. provided the virus, the virus labeled tissues, and helpful discussion. K.C. supervised all aspects of the work.
abstractUnderstanding 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 crosslink with biomolecules. SHIELD preserved 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 (3D) phenotyping of core needle biopsies and human brain cells. SHIELD enables rapid, multiscale, integrated molecular phenotyping of both animal and clinical tissues.
pmid30556815
doi10.1038/nbt.4281
oafree_for_read