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

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216305%3A26620%2F22%3APU143790" target="_blank" >RIV/00216305:26620/22:PU143790 - isvavai.cz</a>

Alternative codes found

RIV/00064165:_____/22:10437367 RIV/00216208:11110/22:10437367

Result on the web

<a href="https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85123910148&origin=inward" target="_blank" >https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85123910148&origin=inward</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1039/d1ra08138a" target="_blank" >10.1039/d1ra08138a</a>

Result language

angličtina

Original language name

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

Original language description

A 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 μL/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.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

20201 - Electrical and electronic engineering

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2022

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

RSC Advances

ISSN

2046-2069

e-ISSN

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Volume of the periodical

12

Issue of the periodical within the volume

4

Country of publishing house

GB - UNITED KINGDOM

Number of pages

8

Pages from-to

2375-2382

UT code for WoS article

000743169000001

EID of the result in the Scopus database

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