Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 61, Issue 20, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202116909
Keywords
Error Correction; Living Cell Imaging; MicroRNA Multiplexed Detection; Nanoparticles; Spatial Resolution
Categories
Funding
- Excellent Young Scientists Fund [22022407]
- National Natural Science Foundation of China [21874008, 22004006]
- Major Program of National Natural Science Foundation of China [21890740, 21890742]
- Special Foundation for State Major Research Program of China [2019YFC1606603]
- SZU Top Ranking Project [860000002100165]
- Beijing Municipal Science and Technology Commission [z131102002813058]
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This study presents a multiplexed miRNA imaging method that utilizes fluorescent label encoding to simultaneously detect multiple miRNAs in individual living cells with error-correcting capability and low cytotoxicity. The method allows accurate quantification and evaluation of breast-cancer-related miRNAs, and provides a single-cell analysis platform.
Simultaneous imaging of multiple microRNAs (miRNAs) in individual living cells is challenging due to the lack of spectrally distinct encoded fluorophores and non-cytotoxic methods. We describe a multiplexed error-robust combinatorial fluorescent label-encoding method, termed fluorophores encoded error-corrected labels (FluoELs), enabling multiplexed miRNA imaging in living cells with error-correcting capability. The FluoELs comprise proportional dual fluorophores for encoding and a constant quantitative single fluorophore for error-corrected quantification. Both are embedded in 260 nm core-shell silica nanoparticles modified with molecular beacon detection probes. The FluoELs are low cytotoxic and could accurately quantify and spatially resolve nine breast-cancer-related miRNAs and evaluate their coordination. The FluoELs enabled a single-cell analysis platform to evaluate miRNA expression profiles and the molecular mechanisms underlying miRNA-associated diseases.
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