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dc.contributor.authorSrinivas, Rathi L.
dc.contributor.authorJohnson, Stephen D.
dc.contributor.authorDoyle, Patrick S.
dc.date.accessioned2016-02-12T17:45:42Z
dc.date.available2016-02-12T17:45:42Z
dc.date.issued2013-11
dc.date.submitted2013-10
dc.identifier.issn0003-2700
dc.identifier.issn1520-6882
dc.identifier.urihttp://hdl.handle.net/1721.1/101171
dc.description.abstractMultiplexed, 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.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.). Center for Future Technologies in Cancer Care (U54-EB-015403-01)en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (Interdepartmental Biotechnology Training Grant T32 GM08334)en_US
dc.description.sponsorshipNational Science Foundation (U.S.) (Grant CMMI-1120724)en_US
dc.language.isoen_US
dc.publisherAmerican Chemical Society (ACS)en_US
dc.relation.isversionofhttp://dx.doi.org/10.1021/ac403201pen_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourcePMCen_US
dc.titleOil-Isolated Hydrogel Microstructures for Sensitive Bioassays On-Chipen_US
dc.typeArticleen_US
dc.identifier.citationSrinivas, Rathi L., Stephen D. Johnson, and Patrick S. Doyle. “Oil-Isolated Hydrogel Microstructures for Sensitive Bioassays On-Chip.” Anal. Chem. 85, no. 24 (December 17, 2013): 12099–12107.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemical Engineeringen_US
dc.contributor.mitauthorSrinivas, Rathi L.en_US
dc.contributor.mitauthorJohnson, Stephen D.en_US
dc.contributor.mitauthorDoyle, Patrick S.en_US
dc.relation.journalAnalytical Chemistryen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsSrinivas, Rathi L.; Johnson, Stephen D.; Doyle, Patrick S.en_US
dc.identifier.orcidhttps://orcid.org/0000-0002-9781-0135
mit.licensePUBLISHER_POLICYen_US


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