Publication detail

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.

Original Title

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

Type

journal article in Web of Science

Language

English

Original Abstract

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.

Keywords

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

Authors

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

Released

30. 3. 2020

Publisher

Springer Nature

ISBN

2045-2322

Periodical

Scientific Reports

Year of study

10

Number

1

State

United Kingdom of Great Britain and Northern Ireland

Pages from

1

Pages to

9

Pages count

9

URL

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

Full text in the Digital Library

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"
}