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Advancing in situ single-cell microbiological analysis through a microwell droplet array with a gradual open sidewall
Jie Wang1, Lin Du2, Yuwei Han3
1School of Optical-Electrical and Computer Engineering, University of Shanghai for Science and Technology, Shanghai, China, 200093. dwzhang@usst.edu.cn.
Lab on a Chip
|November 14, 2023
Summary
This study introduces a novel microwell droplet array chip for rapid, in situ detection of pathogenic bacteria. The technology enables efficient isolation, cultivation, and identification of single bacteria from complex populations.
Area of Science:
- Microfluidics
- Biotechnology
- Analytical Chemistry
Background:
- Microfluidic analysis enhances pathogenic bacteria detection but faces challenges in rapid, in situ single-bacteria array detection due to diffusion limitations.
- Existing methods struggle with complex bacterial populations and low Reynolds number fluids, hindering efficient single-cell analysis.
Purpose of the Study:
- To develop an innovative microwell droplet array chip utilizing lateral hydrodynamic wetting for improved in situ detection of single-bacteria arrays.
- To overcome diffusion limitations and enable rapid, efficient isolation, cultivation, and identification of pathogenic bacteria.
Main Methods:
- Development of microwell droplet array chips with gradually opening sidewalls to facilitate liquid-air interface advancement and microwell impregnation.
- Utilized lateral hydrodynamic wetting to maintain the Wenzel state and resist liquid-air separation.
- Simulated streamlined distribution and performed water-based ink diffusion experiments to assess diffusion efficiency and flow velocity relationships.
Main Results:
- Demonstrated feasibility of preparing and identifying cell arrays within microwells using gradual and traditional designs.
- Smaller gradual microwells showed good diffusion efficiency at 2.1 μL min⁻¹ flow velocity, with easier infiltration state adjustment.
- Successfully isolated mixed populations of E. coli and S. aureus, obtained single-bacteria arrays, and performed in situ Gram assays after propagation.
Conclusions:
- The developed microwell droplet array chip platform effectively addresses challenges in rapid, in situ pathogenic bacteria detection.
- The platform enables successful isolation, cultivation, and detection of single bacteria from mixed populations.
- This technology holds significant potential for advancing diagnostics and research in microbiology and infectious diseases.
