Pneumatic microfluidics-based multiplex single-cell array.

Area of Science:

• Biotechnology
• Microfluidics
• Cell Biology

Background:

• High-throughput screening of cell function and heterogeneity demands large-scale single-cell arrays.
• Existing methods for generating multiplexed, large-scale single-cell arrays are not well-established, hindering rapid and convenient application.

Purpose of the Study:

• To develop a simple, reliable, and universal method for generating large-scale single-cell arrays.
• To demonstrate the capability of multiplexed trapping and concurrent cellular analysis within a single microfluidic device.

Main Methods:

• Integration of pneumatic microvalve arrays (PμVAs) with hydrodynamic single-cell trapping sites in a single microfluidic device.
• Precise control of PμVAs using actuated pressures to guide multiple cell types into designated trapping sites.
• Step-by-step protocol for realizing a multiplexed single-cell array with three distinct cell types, validated by theoretical and computational simulations.

Main Results:

• Successful generation of a multiplexed single-cell array containing three different cell types.
• Concurrent analysis of cellular esterase heterogeneity across the three cell types within the device.
• Demonstration of the method's applicability for generating large-scale single-cell arrays with multiple cell types.

Conclusions:

• The developed microfluidic method offers a promising approach for creating large-scale, multiplexed single-cell arrays.
• This technology has significant potential for applications in high-throughput drug testing, multipurpose immunosensors, and clinical diagnostics.