• Media type: E-Article
  • Title: Rapid Characterization of Biomolecules’ Thermal Stability in a Segmented Flow-Through Optofluidic Microsystem
  • Contributor: Fohlerova, Zdenka; Zhu, Hanliang; Hubalek, Jaromir; Ni, Sheng; Yobas, Levent; Podesva, Pavel; Otahal, Alexandr; Neuzil, Pavel
  • imprint: Springer Science and Business Media LLC, 2020
  • Published in: Scientific Reports
  • Language: English
  • DOI: 10.1038/s41598-020-63620-5
  • ISSN: 2045-2322
  • Keywords: Multidisciplinary
  • Origination:
  • Footnote:
  • Description: <jats:title>Abstract</jats:title><jats:p>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 (∇<jats:italic>T</jats:italic>) 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 (<jats:italic>T</jats:italic><jats:sub>M</jats:sub>) in a flow-through configuration without gradient to verify the functionality of the proposed detection system. We then used the segmented flow configuration and measured the fluorescence amplitude of a droplet exposed to ∇<jats:italic>T</jats:italic> of ≈ 2.31 °C mm<jats:sup>−1</jats:sup>, determining the heat transfer time as ≈ 554 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.</jats:p>
  • Access State: Open Access