Double Emulsion Flow Cytometry for Rapid Single Genome Detection.

Area of Science:

• Biotechnology
• Microfluidics
• Analytical Chemistry

Background:

• Single emulsion (SE) droplet microfluidics enables high-throughput single-cell analysis.
• Double emulsion (DE) droplets offer enhanced stability and compatibility with flow cytometry.

Purpose of the Study:

• To describe a simple, single-layer DE droplet generation device.
• To detail protocols for single-molecule and single-cell assays using DE droplets.
• To facilitate broader adoption of drop-based screening through flow cytometry compatibility.

Main Methods:

• Fabrication of a single-layer DE droplet generation device with plasma treatment for surface wetting control.
• Production of single-core DEs with high monodispersity.
• Application of DE droplets for droplet digital PCR and automated detection on a fluorescence-activated cell sorter (FACS).

Main Results:

• Robust production of monodisperse single-core DEs.
• Successful implementation of single-molecule detection via droplet digital PCR within DEs.
• Automated detection of DE droplets using FACS.

Conclusions:

• The developed DE droplet microfluidics device is simple to fabricate and operate.
• DE droplets are suitable for high-throughput single-molecule and single-cell assays.
• FACS compatibility of DE droplets promotes wider accessibility of drop-based screening technologies.