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"
}