Single cell microRNA analysis using microfluidic flow cytometry.
MicroRNAs (miRNAs) are non-coding small RNAs that have cell type and cell context-dependent expression and function. To study miRNAs at single-cell resolution, we have developed a novel microfluidic approach, where flow fluorescent in situ hybridization (flow-FISH) using locked-nucleic acid probes i...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3559333?pdf=render |
| _version_ | 1818426458594344960 |
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| author | Meiye Wu Matthew Piccini Chung-Yan Koh Kit S Lam Anup K Singh |
| author_facet | Meiye Wu Matthew Piccini Chung-Yan Koh Kit S Lam Anup K Singh |
| author_sort | Meiye Wu |
| collection | DOAJ |
| description | MicroRNAs (miRNAs) are non-coding small RNAs that have cell type and cell context-dependent expression and function. To study miRNAs at single-cell resolution, we have developed a novel microfluidic approach, where flow fluorescent in situ hybridization (flow-FISH) using locked-nucleic acid probes is combined with rolling circle amplification to detect the presence and localization of miRNA. Furthermore, our flow cytometry approach allows analysis of gene-products potentially targeted by miRNA together with the miRNA in the same cells. We demonstrate simultaneous measurement of miR155 and CD69 in 12-O-tetradecanoylphorbol 13-acetate (PMA) and Ionomycin stimulated Jurkat cells. The flow-FISH method can be completed in ∼10 h, utilizes only 170 nL of reagent per experimental condition, and is the first to directly detect miRNA in single cells using flow cytometry. |
| first_indexed | 2024-12-14T14:30:09Z |
| format | Article |
| id | doaj.art-cc120508ae6f40199dea407521a8c19d |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T14:30:09Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-cc120508ae6f40199dea407521a8c19d2022-12-21T22:57:50ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0181e5504410.1371/journal.pone.0055044Single cell microRNA analysis using microfluidic flow cytometry.Meiye WuMatthew PicciniChung-Yan KohKit S LamAnup K SinghMicroRNAs (miRNAs) are non-coding small RNAs that have cell type and cell context-dependent expression and function. To study miRNAs at single-cell resolution, we have developed a novel microfluidic approach, where flow fluorescent in situ hybridization (flow-FISH) using locked-nucleic acid probes is combined with rolling circle amplification to detect the presence and localization of miRNA. Furthermore, our flow cytometry approach allows analysis of gene-products potentially targeted by miRNA together with the miRNA in the same cells. We demonstrate simultaneous measurement of miR155 and CD69 in 12-O-tetradecanoylphorbol 13-acetate (PMA) and Ionomycin stimulated Jurkat cells. The flow-FISH method can be completed in ∼10 h, utilizes only 170 nL of reagent per experimental condition, and is the first to directly detect miRNA in single cells using flow cytometry.http://europepmc.org/articles/PMC3559333?pdf=render |
| spellingShingle | Meiye Wu Matthew Piccini Chung-Yan Koh Kit S Lam Anup K Singh Single cell microRNA analysis using microfluidic flow cytometry. PLoS ONE |
| title | Single cell microRNA analysis using microfluidic flow cytometry. |
| title_full | Single cell microRNA analysis using microfluidic flow cytometry. |
| title_fullStr | Single cell microRNA analysis using microfluidic flow cytometry. |
| title_full_unstemmed | Single cell microRNA analysis using microfluidic flow cytometry. |
| title_short | Single cell microRNA analysis using microfluidic flow cytometry. |
| title_sort | single cell microrna analysis using microfluidic flow cytometry |
| url | http://europepmc.org/articles/PMC3559333?pdf=render |
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