Gradient Nanostructures and Machine Learning Synergy for Robust Quantitative Surface-Enhanced Raman Scattering

Surface-Enhanced Raman Scattering (SERS) holds significant promise for trace-level molecular detection but faces challenges in achieving reliable quantitative analysis due to signal variability caused by non-uniform "hot spots" and external factors. To address these limitations, a novel SERS platform based on gradient nanostructures is developed using shadow sphere lithography, enabling the acquisition of diverse spectral features from a single analyte concentration under identical conditions. The gradient design minimizes fabrication variability and enhances spectral diversity, while the machine learning (ML) model trained on the multi-spectral dataset significantly outperformed traditional single-spectrum approaches, with the test Mean Squared Error (MSE) reduced by 84.8% and the coefficient of determination (R²) improved by 61.2%. This strategy captures subtle spectral variations, improving the precision, robustness, and reproducibility of SERS-based quantification, paving the way for its reliable application in real-world scenarios.