Detail publikace
Rapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystem
Originální název
Rapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystem
Anglický název
Rapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystem
Jazyk
en
Originální abstrakt
Optofluidic 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.
Anglický abstrakt
Optofluidic 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.
Plný text v Digitální knihovně
Dokumenty
BibTex
@article{BUT163206,
author="Zdenka {Fohlerová} and Hanliang {Zhu} and Jaromír {Hubálek} and Sheng {Ni} and Pavel {Podešva} and Alexandr {Otáhal} and Pavel {Neužil} and Levent {Yobas}",
title="Rapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystem",
annote="Optofluidic 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.",
address="Springer Nature",
chapter="163206",
doi="10.1038/s41598-020-63620-5",
howpublished="online",
institution="Springer Nature",
number="1",
volume="10",
year="2020",
month="march",
pages="1--7",
publisher="Springer Nature",
type="journal article in Web of Science"
}