Dataset Information

Sensitive and multiplexed on-chip microRNA profiling in oil-isolated hydrogel chambers.

ABSTRACT: Although microRNAs (miRNAs) have been shown to be excellent indicators of disease state, current profiling platforms are insufficient for clinical translation. Here, we demonstrate a versatile hydrogel-based microfluidic approach and novel amplification scheme for entirely on-chip, sensitive, and highly specific miRNA detection without the risk of sequence bias. A simulation-driven approach is used to engineer the hydrogel geometry and the gel-reaction environment is chemically optimized for robust detection performance. The assay provides 22.6?fM sensitivity over a three log range, demonstrates multiplexing across at least four targets, and requires just 10.3?ng of total RNA input in a 2?hour and 15?minutes assay.

SUBMITTER: Lee H

PROVIDER: S-EPMC4547856 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Sensitive and multiplexed on-chip microRNA profiling in oil-isolated hydrogel chambers.

Lee Hyewon H Srinivas Rathi L RL Gupta Ankur A Doyle Patrick S PS

Angewandte Chemie (International ed. in English) 20150107 8

Although microRNAs (miRNAs) have been shown to be excellent indicators of disease state, current profiling platforms are insufficient for clinical translation. Here, we demonstrate a versatile hydrogel-based microfluidic approach and novel amplification scheme for entirely on-chip, sensitive, and highly specific miRNA detection without the risk of sequence bias. A simulation-driven approach is used to engineer the hydrogel geometry and the gel-reaction environment is chemically optimized for rob ...[more]

PMID: 25565630

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Project description:Multiplexed, sensitive, and on-chip molecular diagnostic assays are essential in both clinical and research settings. In past work, running reactions in nanoliter- to femtoliter-sized volumes such as microwells or droplets has led to significant increases in detection sensitivities. At the same time, hydrogels have emerged as attractive scaffolds for bioassays due to their nonfouling, flexible, and aqueous properties. In this paper, we combine these concepts and develop a novel platform in which hydrogel compartments are used as individually confined reaction volumes within a fluorinated oil phase. We fabricate functional and versatile hydrogel microstructures in microfluidic channels that are physically isolated from each other using a surfactant-free fluorinated oil phase, generating picoliter- to nanoliter-sized immobilized aqueous reaction compartments that are readily functionalized with biomolecules. In doing so, we achieve monodisperse reaction volumes with an aqueous interior while exploiting the unique chemistry of a hydrogel, which provides a solid and porous binding scaffold for biomolecules and is impenetrable to oil. Furthermore, our lithographically defined reaction volumes are readily customized with respect to geometry and chemistry within the same channel, allowing rational tuning of the confined reaction volume on a post-to-post basis without needing to use surfactants to maintain stability. We design and implement a multiplexed signal amplification assay in which gel-bound enzymes turn over small molecule substrate into fluorescent product in the oil-confined gel compartment, providing significant signal enhancement. Using short (20 min) amplification times, the encapsulation scheme provides up to 2 orders of magnitude boost of signal in nucleic acid detection assays relative to direct labeling and does not suffer from any cross-talk between the posts. We ultimately demonstrate up to 57-fold increase in nucleic acid detection sensitivity compared to a direct labeling scheme.

Dataset Information

Sensitive and multiplexed on-chip microRNA profiling in oil-isolated hydrogel chambers.

Publications

Sensitive and multiplexed on-chip microRNA profiling in oil-isolated hydrogel chambers.

Similar Datasets

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