Rapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystem

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dc.contributor.authorFohlerová, Zdenkacs
dc.contributor.authorHubálek, Jaromírcs
dc.contributor.authorPodešva, Pavelcs
dc.contributor.authorOtáhal, Alexandrcs
dc.contributor.authorNeužil, Pavelcs
dc.coverage.issue1cs
dc.coverage.volume10cs
dc.date.accessioned2020-08-04T11:00:37Z
dc.date.available2020-08-04T11:00:37Z
dc.date.issued2020-03-30cs
dc.description.abstractOptofluidic devices combining optics and microfluidics have recently attracted attention for biomolecular analysis due to their high detection sensitivity. Here, we show a silicon chip with tubular microchannels buried inside the substrate featuring temperature gradient (T) along the microchannel. We set up an optical fluorescence system consisting of a power-modulated laser light source of 470 nm coupled to the microchannel serving as a light guide via optical fiber. Fluorescence was detected on the other side of the microchannel using a photomultiplier tube connected to an optical fiber via a fluorescein isothiocyanate filter. The PMT output was connected to a lock-in amplifier for signal processing. We performed a melting curve analysis of a short dsDNA – SYBR Green I complex with a known melting temperature (TM) in a flow-through configuration without gradient to verify the functionality of proposed detection system. We then used the segmented flow configuration and measured the fluorescence amplitude of a droplet exposed to T of 2.31°C mm-1, determining the heat transfer time as 563 ms. The proposed platform can be used as a fast and cost-effective system for performing either MCA of dsDNAs or for measuring protein unfolding for drug-screening applications.en
dc.formattextcs
dc.format.extent1-9cs
dc.format.mimetypeapplication/pdfcs
dc.identifier.citationScientific Reports. 2020, vol. 10, issue 1, p. 1-9.en
dc.identifier.doi10.1038/s41598-020-63620-5cs
dc.identifier.issn2045-2322cs
dc.identifier.other163206cs
dc.identifier.urihttp://hdl.handle.net/11012/193484
dc.language.isoencs
dc.publisherSpringer Naturecs
dc.relation.ispartofScientific Reportscs
dc.relation.urihttps://www.nature.com/articles/s41598-020-63620-5cs
dc.rightsCreative Commons Attribution 4.0 Internationalcs
dc.rights.accessopenAccesscs
dc.rights.sherpahttp://www.sherpa.ac.uk/romeo/issn/2045-2322/cs
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/cs
dc.subjectoptofluidicsen
dc.subjecttemperature gradienten
dc.subjectmelting temperature of dsDNAen
dc.subjectprotein unfoldingen
dc.titleRapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystemen
dc.type.driverarticleen
dc.type.statusPeer-revieweden
dc.type.versionpublishedVersionen
sync.item.dbidVAV-163206en
sync.item.dbtypeVAVen
sync.item.insts2021.02.25 16:53:16en
sync.item.modts2021.02.25 16:13:29en
thesis.grantorVysoké učení technické v Brně. Fakulta elektrotechniky a komunikačních technologií. Ústav mikroelektronikycs
thesis.grantorVysoké učení technické v Brně. Středoevropský technologický institut VUT. Chytré nanonástrojecs
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