Temperature non-uniformity detection on dPCR chips and temperature sensor calibration

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dc.contributor.authorGaňová, Martinacs
dc.contributor.authorWang, Xinlucs
dc.contributor.authorYan, Zhiqiangcs
dc.contributor.authorZhang, Haoqingcs
dc.contributor.authorLednický, Tomášcs
dc.contributor.authorKorabečná, Mariecs
dc.contributor.authorNeužil, Pavelcs
dc.coverage.issue4cs
dc.coverage.volume12cs
dc.date.accessioned2022-05-31T14:55:38Z
dc.date.available2022-05-31T14:55:38Z
dc.date.issued2022-01-12cs
dc.description.abstractA microfluidic-based digital polymerase chain reaction (dPCR) chip requires precise temperature control as well as uniform temperature distribution to ensure PCR efficiency. However, measuring local temperature and its distribution over thousands of L/nL-volume samples with minimum disturbance is challenging. Here, we present a method of non-contact localized temperature measurement for determination of the non-uniformity of temperature distribution over a dPCR chip. We filled the dPCR chip with a PCR solution containing amplified DNA fragments with a known melting temperature (TM). We then captured fluorescent images of the chip when it was heated from 70 to 99 °C, plotted the fluorescence intensity of each partition as a function of temperature, and calculated measured TM values from each partition. Finally, we created a 3-D map of the dPCR chip with the measured TM as the parameter. Even when the actual TM of the PCR solution was constant, the measured TM value varied between locations due to temperature non-uniformity in the dPCR chip. The method described here thereby characterized the distribution of temperature non-uniformity using a PCR solution with known TM as a temperature sensor. Among the non-contact temperature measurement methods, the proposed TM-based method can determine the temperature distribution within the chip, instead of only at the chip surface. The method also does not suffer from the undesirable photobleaching effect of fluorescein-based temperature measurement method. Temperature determination over the dPCR chip based on TM allowed us to calibrate the temperature sensor and improve the dPCR configuration and precision. This method is also suitable for determining the temperature uniformity of other microarray systems where there is no physical access to the system and thus direct temperature measurement is not possible.en
dc.formattextcs
dc.format.extent2375-2382cs
dc.format.mimetypeapplication/pdfcs
dc.identifier.citationRSC Advances. 2022, vol. 12, issue 4, p. 2375-2382.en
dc.identifier.doi10.1039/d1ra08138acs
dc.identifier.issn2046-2069cs
dc.identifier.other176929cs
dc.identifier.urihttp://hdl.handle.net/11012/204605
dc.language.isoencs
dc.publisherRoyal Society of Chemistrycs
dc.relation.ispartofRSC Advancescs
dc.relation.urihttps://pubs.rsc.org/en/content/articlelanding/2022/RA/D1RA08138Acs
dc.rightsCreative Commons Attribution-NonCommercial 4.0 Internationalcs
dc.rights.accessopenAccesscs
dc.rights.sherpahttp://www.sherpa.ac.uk/romeo/issn/2046-2069/cs
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/cs
dc.subjectFluorescenceen
dc.subjectPhotobleachingen
dc.subjectPolymerase chain reactionen
dc.subjectTemperature measurementen
dc.subjectTemperature sensorsen
dc.subjectThermometersen
dc.titleTemperature non-uniformity detection on dPCR chips and temperature sensor calibrationen
dc.type.driverarticleen
dc.type.statusPeer-revieweden
dc.type.versionpublishedVersionen
sync.item.dbidVAV-176929en
sync.item.dbtypeVAVen
sync.item.insts2022.06.22 00:54:28en
sync.item.modts2022.06.22 00:14:24en
thesis.grantorVysoké učení technické v Brně. Středoevropský technologický institut VUT. Chytré nanonástrojecs
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