Detail publikace

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

FOHLEROVÁ, Z. ZHU, H. HUBÁLEK, J. NI, S. PODEŠVA, P. OTÁHAL, A. NEUŽIL, P. YOBAS, L.

Originální název

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

Typ

článek v časopise ve Web of Science, Jimp

Jazyk

angličtina

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.

Klíčová slova

optofluidics, temperature gradient, melting temperature of dsDNA, protein unfolding

Autoři

FOHLEROVÁ, Z.; ZHU, H.; HUBÁLEK, J.; NI, S.; PODEŠVA, P.; OTÁHAL, A.; NEUŽIL, P.; YOBAS, L.

Vydáno

30. 3. 2020

Nakladatel

Springer Nature

ISSN

2045-2322

Periodikum

Scientific Reports

Ročník

10

Číslo

1

Stát

Spojené království Velké Británie a Severního Irska

Strany od

1

Strany do

9

Strany počet

9

URL

https://www.nature.com/articles/s41598-020-63620-5

Plný text v Digitální knihovně

http://hdl.handle.net/11012/193484

BibTex

@article{BUT163206,
  author="FOHLEROVÁ, Z. and ZHU, H. and HUBÁLEK, J. and NI, S. and PODEŠVA, P. and OTÁHAL, A. and NEUŽIL, P. and YOBAS, L.",
  title="Rapid Characterization of Biomolecules’ Thermal Stability in Segmented Flow-Through Optofluidic Microsystem",
  journal="Scientific Reports",
  year="2020",
  volume="10",
  number="1",
  pages="1--9",
  doi="10.1038/s41598-020-63620-5",
  issn="2045-2322",
  url="https://www.nature.com/articles/s41598-020-63620-5"
}