Wachara Chanakul, Nattapong Chantipmanee, Matthieu Sandell, Ruoming An, Michael Mayer, Yan Xu
Nanopore technologies have become powerful tools for single-molecule detection and sequencing, offering unprecedented insight into molecular structures and dynamics. This review summarizes key advances in nanopore fabrication, surface modification, and integration with chip-based nanofluidic devices. Such integration enhances analytical performance by enabling precise molecular control, reducing electrical noise, improving reproducibility, and facilitating automated handling. Despite substantial progress, challenges remain, including accurate control of pore geometry, fabrication reproducibility, material stability, nonspecific interactions, and scaling for high-throughput analysis. Integrating nanopores with nanofluidic systems provides promising routes to overcome these barriers, while emerging applications of artificial intelligence and advanced data analysis further enhance signal interpretation and sensitivity. By assessing recent progress and persistent challenges, this review highlights future directions toward robust, scalable nanopore–nanofluidic platforms that could transform analytical chemistry, enabling breakthroughs in biomolecular sensing, diagnostics, environmental monitoring, and personalized medicine.