A Solid-State Hard Microfluidic-Nanopore Biosensor with Multilayer Fluidics and On-Chip Bioassay/Purification Chamber

Area of Science:

• Biotechnology
• Nanotechnology
• Analytical Chemistry

Background:

• Solid-state nanopore sensors offer potential for nucleic acid and protein analysis.
• Clinical application of nanopore sensing is hindered by complex sample preparation and fluid handling requirements.
• Current setups require multiple components, limiting point-of-care diagnostic translation.

Purpose of the Study:

• To develop an integrated microfluidic device combining bioassay reaction/purification with a solid-state nanopore sensor.
• To overcome limitations of conventional nanopore setups for clinical and point-of-care applications.
• To demonstrate the utility of the device for on-chip single-molecule DNA detection.

Main Methods:

• Fabrication of a microfluidic device using Zeonex plastic with micro-machining and heat bonding.
• Integration of a heating element, magnetic controller, and fluid port selector valves for precise fluid control.
• Performance evaluation through electrical noise measurements and DNA translocation experiments.
• Demonstration of on-chip DNA Polymerase Chain Reaction (PCR) product detection post-magnetic bead separation.

Main Results:

• The microfluidic-nanopore device demonstrated comparable performance to conventional setups.
• The integrated system significantly improved ease of use compared to traditional nanopore sensors.
• Successful single-molecule detection of DNA PCR products was achieved on-chip.
• The device exhibited high-temperature and solvent resistance, enabling versatile applications.

Conclusions:

• The developed microfluidic-nanopore device represents a significant advancement for biosensing.
• The integrated design facilitates sample preparation, purification, and detection in a single unit.
• This technology shows promise for enabling portable and user-friendly point-of-care nucleic acid diagnostics.