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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
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Related Experiment Video

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Author Spotlight: Characterization of Low-Affinity Protein Interactions in Solution Using MassFluidix Technology
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Spatially Resolved Protein Binding Kinetics Analysis in Microfluidic Photonic Crystal Sensors.

Stefanie Lehmann1, Fabio Aldo Kraft1, Martina Gerken1

  • 1Integrated Systems and Photonics, Faculty of Engineering, Kiel University, 24118 Kiel, Germany.

Sensors (Basel, Switzerland)
|July 8, 2023
PubMed
Summary

This study introduces photonic crystal slabs for label-free biomarker detection in Organ-on-a-Chip systems. The method enables non-contact monitoring of binding kinetics within microfluidic devices, advancing drug screening and medical research.

Keywords:
diffusionin-channel referencinglab-on-a-chiplabel-free biosensormicrofluidicsorgan-on-a-chipphotonic crystal biosensorprotein binding kineticsspatially resolved spectral analysis

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Area of Science:

  • Biophotonics
  • Microfluidics
  • Biomolecular Engineering

Background:

  • Organ-on-a-Chip systems are crucial for in vitro drug screening and medical research.
  • Label-free, continuous biomolecular monitoring is needed for cell culture analysis in microfluidic systems.
  • Photonic crystal slabs offer a promising optical transduction method for label-free detection.

Purpose of the Study:

  • To investigate the use of photonic crystal slabs integrated with microfluidic chips for label-free biomarker detection.
  • To analyze the capability of same-channel reference for protein binding measurements using a spectrometer and 1D spatially resolved data.
  • To evaluate the performance of a cross-correlation-based data analysis for binding kinetics.

Main Methods:

  • Integration of photonic crystal slabs with microfluidic chips.
  • Label-free detection using optical spectroscopy and 1D spatial resolution (1.2 μm).
  • Implementation of a cross-correlation-based data analysis procedure.
  • Determination of limit of detection (LOD) using an ethanol-water dilution series.
  • Assessment of binding kinetics using a streptavidin-biotin interaction model.

Main Results:

  • The median LOD was determined to be (2.3±0.4)×10-4 RIU (10s exposure) and (1.3±0.24)×10-4 RIU (30s exposure).
  • Localized protein binding was achieved within the microfluidic channel under laminar flow conditions.
  • Binding kinetics were observed to diminish at the microfluidic channel edges due to the velocity profile.

Conclusions:

  • Photonic crystal slabs integrated with microfluidics provide a viable platform for label-free biomarker detection.
  • The developed system allows for non-contact monitoring of binding kinetics with spatial resolution.
  • Understanding flow dynamics is crucial for optimizing biomolecular detection at microfluidic channel edges.