With the growing popularity of the non-destructive technique, surface-enhanced Raman spectroscopy (SERS) demands a highly sensitive and reproducible plasmonic nanoparticles substrate. In this study, a novel bimetallic core-shell nanoparticles (Au@Ag@mSiO₂NP) substrate consisting of a gold core, silver shell, and a mesoporous silica coating was synthesized. The mesoporous coating structure was created by employing template molecules such as surfactant and their subsequent removal allowing selective screening based on the size of analyte molecules. Results showed that the plasmonic substrate could selectively enhance small molecules by preventing large macromolecules to reach the exciting zone of the substrate core, achieving the detection of chloramphenicol in milk samples with a detection limit of 6.68 × 10^-8 M. Moreover, the mesoporous coating provided additional stability to the Au@Ag nanoparticles, leading to the reusability of the substrate. Thus, this work offered a simple and smart Au@Ag@mSiO₂NP substrate for effective SERS detection of analytes.

The conventional microbial cell analyses are mostly population-averaged methods that conceal the characteristics of single-cell in the community. Single-cell analysis can provide information on the functional and structural variation of each cell, resulting in the elimination of long and tedious microbial cultivation techniques. Raman spectroscopy is a label-free, noninvasive, and in-vivo method ideal for single-cell measurement to obtain spatially resolved chemical information. In the current review, recent developments in Raman spectroscopic techniques for microbial characterization at the single-cell level are presented, focusing on Raman imaging of single cells to study the intracellular distribution of different components. The review also discusses the limitation and challenges of each technique and put forward some future outlook for improving Raman spectroscopy-based techniques for single-cell analysis. Raman spectroscopic methods at the single-cell level have potential in precision measurements, metabolic analysis, antibiotic susceptibility testing, resuscitation capability, and correlating phenotypic information to genomics for cells, the integration of Raman spectroscopy with other techniques such as microfluidics, stable isotope probing (SIP), and atomic force microscope can further improve the resolution and provide extensive information. Future focuses should be given to advance algorithms for data analysis, standardized reference libraries, and automated cell isolation techniques in future.