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Single-cell activity screening in microfluidic droplets.

Stefanie Neun1, Tomasz S Kaminski1, Florian Hollfelder1

  • 1Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom.

Methods in Enzymology
|November 1, 2019
PubMed
Summary
This summary is machine-generated.

Single cells are encapsulated in water-in-oil emulsion droplets for activity assays. Microfluidic devices enable ultrahigh-throughput screening using optical detection methods.

Keywords:
Directed evolutionDroplet microfluidicsFunctional metagenomics

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Cell Biology

Background:

  • Water-in-oil emulsion droplets serve as microcompartments for single-cell analysis.
  • Microfluidic technology enables high-throughput droplet generation and manipulation.

Purpose of the Study:

  • To describe the use of microfluidic droplets for cell-based activity assays.
  • To highlight the capabilities of optical detection within these microcompartments.
  • To demonstrate applications in directed evolution and functional metagenomics.

Main Methods:

  • Generating water-in-oil emulsion droplets using microfluidic devices at high rates (> kHz).
  • Coupling microfluidic devices with modules for reagent addition, incubation, and optical measurements.
  • Employing fluorescence and absorbance detection for analyte concentration measurement (down to 2nM sensitivity).

Main Results:

  • Demonstration of single-cell encapsulation and activity assays within microfluidic droplets.
  • Achieved high-throughput optical detection with low analyte sensitivity.
  • Successful application of the technology for ultrahigh-throughput sorting.

Conclusions:

  • Microfluidic droplet technology provides a powerful platform for high-throughput single-cell analysis.
  • Optical detection methods integrated with microfluidics enable sensitive and efficient screening.
  • This approach is valuable for advancing directed evolution and functional metagenomics research.