Rapid detection and quantification of ultralow-concentration nanoparticles based on nanopore array blockage

Abstract

Rapid and accurate detection of ultralow-concentration nanoparticles is crucial for applications ranging from medical diagnosis to water quality monitoring, yet remains challenging for current laser-based and light-scattering methods. While nanoparticle-translocation-based nanopore sensing offers single-particle resolution, conventional single-nanopore resistive pulse sensing approaches suffer from low capture frequency, transient signals, and clogging issues, limiting their effectiveness at extremely low concentrations. Here, we present a novel nanopore array blockage-based sensing strategy for the rapid detection and quantification of ultralow-concentration nanoparticles. Using hydraulic force, nanoparticles are driven through an array of subnanoparticle-sized pores, and optical microscopy monitors blockage progression to obtain quantitative concentration data. Our results demonstrate a linear correlation between the initial blockage rate and nanoparticle concentration, enabling the detection of fluorescent nanoparticles down to 0.5 aM (300 particles/mL) within 5 min—a three-order-of-magnitude improvement in sensitivity over previous nanopore-based methods. Additionally, our approach can leverage fluorescent nanoparticles as probes to detect unlabeled nanoparticles and contaminants at similarly low concentrations. This strategy provides a robust, efficient, and rapid platform for ultrasensitive nanoparticle detection, with promising applications in biomedical research, environmental monitoring, and industrial quality control.

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2025 The Author(s). Droplet published by Jilin University and John Wiley & Sons Australia, Ltd.