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High-Density Microfluidic Chip with Vertical Structure for Digital PCR.

Peng Sun¹, Huaqing Si², Gangwei Xu³

¹School of Information Technology, Luoyang Normal University, Luoyang 471934, China.

Sensors (Basel, Switzerland)

|September 13, 2025

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We developed a novel microfluidic chip for digital PCR (polymerase chain reaction) that enhances sensitivity and dynamic range. This low-cost, vertical-structured chip improves nucleic acid quantification for applications like cancer screening.

Area of Science:

Biotechnology
Molecular Biology
Microfluidics

Background:

Digital PCR (polymerase chain reaction) is crucial for absolute nucleic acid quantification in fields like cancer screening.
Current digital PCR methods face limitations including high costs, complex procedures, and restricted detection dynamic range.

Purpose of the Study:

To develop a cost-effective, high-performance microfluidic chip for digital PCR.
To overcome the limitations of existing digital PCR technologies through an innovative vertical structure.

Main Methods:

A novel three-layer glass-PDMS-glass microfluidic chip was designed with a high-density vertical structure.
The chip incorporates 30,000 reaction chambers (0.713 nL each) within the polydimethylsiloxane (PDMS) layer.
The vertical design increases chamber numbers and total volume by 50% without altering chip footprint or chamber size.

Keywords:

detection accuracy digital PCR dynamic range microfluidic chip vertical structure

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Main Results:

The vertical-structured chip theoretically achieves a nucleic acid detection dynamic range approaching 10⁵.
Experiments with serially diluted KRAS plasmid DNA confirmed the chip's accuracy and reliability in quantitative detection.
The chip demonstrated enhanced sensitivity and a significantly wider detection dynamic range compared to conventional designs.

Conclusions:

The developed vertical-structured digital PCR chip offers a simple manufacturing process, uniform sample partitioning, and broad dynamic range.
This innovation has the potential to reduce costs and complexity, promoting wider adoption of digital PCR in biomedical applications.