Electrocontrolled Injection-Coupled Droplet Microfluidic Platform for Antimicrobial Resistance Screening.

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Biology

Background:

• Droplet microfluidics offers high-throughput single-cell analysis.
• Current methods face challenges in controlling single-bacterium encapsulation for antibiotic susceptibility testing (AST).

Purpose of the Study:

• To develop an integrated droplet microfluidic platform for precise single-bacterium encapsulation and rapid AST.
• To enable efficient intradroplet reagent delivery and dual-fluorescence monitoring for bacterial viability and metabolic activity.

Main Methods:

• Electrocontrolled injection for precise bacterial encapsulation.
• Optimization of flow parameters for monodisperse droplet generation and stability.
• Pressure-volume calibration and electroinjection threshold for accurate volume regulation.
• Coencapsulation of Escherichia coli, pGLO plasmid, and resazurin for dual-fluorescence monitoring (GFP and resorufin).

Main Results:

• Achieved controllable single-bacterium encapsulation.
• Demonstrated highly monodisperse droplets with stable morphology.
• Completed AST within 3 hours via fluorescence readout.
• Realized dual-fluorescence monitoring indicating bacterial viability and metabolic activity.

Conclusions:

• The developed multichannel droplet microfluidic system provides a robust and efficient tool for rapid antimicrobial resistance detection.
• The platform integrates miniaturization, high throughput, and low reagent consumption.
• Shows considerable potential for clinical translation in AST.