Extreme sensitivity label-free biosensing platform based on topologically disruptive phase nano-optics

Label-free plasmonic biosensors hold significant promise for advancing molecular biology research and enabling early disease diagnostics, particularly in detecting trace amounts of target molecules in highly diluted solutions. However, conventional plasmonic biosensors face substantial challenges in detecting lower-molecule-weight (<500 Da) biomolecules, especially at extreme low concentrations. To overcome this limitation, a novel plasmonic biosensor platform based on enhanced topological phase singularity has been proposed. Herein, we demonstrate that extremely small-sized (<3 nm) silver nanoparticles, embedded in aluminum oxide with a highly ordered configuration and sub-nanometer alignment and inter-particle spacing (<1 nm), and covered by a gold film, exhibit a topologically dark reflection and produces a sharp phase change, which ultimately presents as a significant shift in the reflected beam's position. By precisely modulating the concentration of silver nanoparticles, a largest position shift of 554.3 µm was achieved in calibration experiments with an extreme sensitivity of 3.27 × 10⁸nm RIU^-1 (refractive index unit). The refractive index resolution was determined to be 3.67 × 10^-7 RIU with a 0.12 µm position resolution. We also demonstrated the ability of the proposed biosensor platform to detect extremely low-molecular-weight (244 Da) biotin biomolecules at femtomolar concentrations. Furthermore, in target biosensing experiments, the plasmonic biosensor platform successfully directly detected small cytokine biomarker (TNF-α) in real-time at 100 zeptomolar (10^-21mol L^-1), highlighting its potential for extreme-sensitive biosensing applications.