Three‐dimensional composite substrate based on pyramidal pitted silicon array adhered Au@Ag nanospheres for high‐performance surface‐enhanced Raman scattering

As a noninvasive and label-free optical technique, Raman spectroscopy offers significant advantages in studying the structure and properties of biomacromolecules, as well as real-time changes in cellular molecular structure. However, its practical applications are hindered by weak scattering responses, low signal intensity, and poor spectral uniformity, which affect the subsequent accuracy of spectral analysis. To address these issues, we report a novel surface-enhanced Raman scattering (SERS) substrate based on a pyramidal pitted silicon (PPSi) array structure adhered with Au-shell Ag-core nanospheres (Au@Ag NSs). By preparing a highly uniform PPSi array substrate with controllable size and arrangement, and constructing SERS-active Au@Ag NSs on this substrate, a three-dimensional (3D) composite SERS substrate is realized. The enhancement performance and spectral uniformity of 3D composite SERS substrate were examined using crystal violet (CV) and Rhodamine 6G (R6G) molecules, achieving a minimum detectable concentration of R6G at 10^-9 M and the analytical enhancement factor (AEF) of 4.2 × 10⁸. Moreover, SERS detection of biological samples with varying concentrations of <i>Staphylococcus aureus</i> demonstrated excellent biocompatibility of the SERS substrate and enabled quantitative analysis of bacterial concentration (<i>R</i> ² = 99.7 %). Theoretical simulations using finite-difference time-domain (FDTD) analysis were conducted to examine the electromagnetic field distribution of the three-dimensional SERS composite substrate, confirming its local electric field enhancement effect. These experimental and theoretical results indicate that the Au@Ag NSs/PPSi substrate with a regulable pyramidal pitted array is a promising candidate for sensitive, label-free SERS detection in medical and biotechnological applications.