Protection of tissue physicochemical properties using polyfunctional crosslinkers.

Nature biotechnology (2018-12-18)
Young-Gyun Park, Chang Ho Sohn, Ritchie Chen, Margaret McCue, Dae Hee Yun, Gabrielle T Drummond, Taeyun Ku, Nicholas B Evans, Hayeon Caitlyn Oak, Wendy Trieu, Heejin Choi, Xin Jin, Varoth Lilascharoen, Ji Wang, Matthias C Truttmann, Helena W Qi, Hidde L Ploegh, Todd R Golub, Shih-Chi Chen, Matthew P Frosch, Heather J Kulik, Byung Kook Lim, Kwanghun Chung

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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Dextran sulfate sodium salt from Leuconostoc spp., for molecular biology, average Mw >500,000 (dextran starting material), contains 0.5-2% phosphate buffer
Sodium acrylate, 97%
Deoxyribonucleic acid, single stranded from salmon testes, For hybridization
Acrylamide, suitable for electrophoresis, ≥99% (HPLC), powder
Trimethylamine N-oxide, 95%
D.E.R. 332, used as embedding medium
(R)-(−)-Glycidyl methyl ether, 97%
Anti-RLN3 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution

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