A novel dual-well array chip for efficiently trapping single-cell in large isolated micro-well without complicated...

Area of Science:

• Biotechnology
• Microfluidics
• Cell Biology

Background:

• Conventional cell-sized well arrays offer high occupancy but lack space for reagents, limiting their use in single-cell analysis.
• This restricts the application of well arrays as standalone single-cell research tools.

Purpose of the Study:

• To develop a novel dual-well array chip integrating capture and reaction wells for enhanced single-cell analysis.
• To establish a convenient flow method for single-cell analysis with high efficiency and low cost.

Main Methods:

• A dual-well chip design with 20 µm capture-wells and 100 µm reaction-wells was developed.
• Cells were loaded by gravity, followed by washing out multi-cells and reagent loading via scraping.
• Computational simulations were used to understand the dual-well structure's working principle.

Main Results:

• Achieved highly efficient single-cell trapping (75.8%) with a low multi-cell rate (11.3%).
• Successfully measured intracellular β-galactosidase activity in K562 cells at the single-cell level.
• Identified a relationship between wall shear stress and aspect ratio for dual-well chip design.

Conclusions:

• The dual-well chip is the first to offer high single-cell occupancy and sufficient reagent space, with low cost and simple operation.
• The developed method provides a feasible and convenient tool for practical single-cell research.
• Computational guidance aids in designing future multi-well chips for single-cell analysis.