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...

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Main Authors:	Wu, Meiye, Piccini, Matthew, Koh, Chung-Yan, Lam, Kit S., Singh, Anup K.
Format:	Online
Language:	English
Published:	Public Library of Science 2013
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3559333/

id	pubmed-3559333
recordtype	oai_dc
spelling	pubmed-35593332013-02-04 Single Cell MicroRNA Analysis Using Microfluidic Flow Cytometry Wu, Meiye Piccini, Matthew Koh, Chung-Yan Lam, Kit S. Singh, Anup K. Research Article 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. Public Library of Science 2013-01-30 /pmc/articles/PMC3559333/ /pubmed/23383050 http://dx.doi.org/10.1371/journal.pone.0055044 Text en © 2013 Wu et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Wu, Meiye Piccini, Matthew Koh, Chung-Yan Lam, Kit S. Singh, Anup K.
spellingShingle	Wu, Meiye Piccini, Matthew Koh, Chung-Yan Lam, Kit S. Singh, Anup K. Single Cell MicroRNA Analysis Using Microfluidic Flow Cytometry
author_facet	Wu, Meiye Piccini, Matthew Koh, Chung-Yan Lam, Kit S. Singh, Anup K.
author_sort	Wu, Meiye
title	Single Cell MicroRNA Analysis Using Microfluidic Flow Cytometry
title_short	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_sort	single cell microrna analysis using microfluidic flow cytometry
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.
publisher	Public Library of Science
publishDate	2013
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3559333/
_version_	1611950871108648960

Single Cell MicroRNA Analysis Using Microfluidic Flow Cytometry

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