Surface-enhanced Raman spectroscopy (SERS) sensing of DNA bases by plasmonic nanopores could pave a way to novel methods for DNA analyses and new generation single-molecule sequencing platforms. The SERS discrimination of single DNA bases depends critically on the time that a DNA strand resides within the plasmonic hot spot. In fact, DNA molecules flow through the nanopores so rapidly that the SERS signals collected are not sufficient for single-molecule analysis. Here, we report an approach to control the residence time of molecules in the hot spot by an electro-plasmonic trapping effect. By directly adsorbing molecules onto a gold nanoparticle and then trapping the single nanoparticle in a plasmonic nanohole up to several minutes, we demonstrate single-molecule SERS detection of all four DNA bases as well as discrimination of single nucleobases in a single oligonucleotide. Our method can be extended easily to label-free sensing of single-molecule amino acids and proteins.

Surface‐enhanced Raman spectroscopy (SERS) has been combined with microfluidic Lab‐on‐a‐Chip (LoC) systems for sensitive optofluidic detection for more than a decade. However, most microfluidic SERS devices still suffer from analyte contamination and signal irreproducibility. In recent years, both the microfluidics and SERS communities have developed their own solutions that are complementary to each other; their combination even has potential for commercialization. In this review, the recent advances in both fields are summarized with regard to the development of reliable multifunctional SERS‐enabled LoC systems and their broad applications. Starting from SERS fundamentals, reproducible SERS substrates and dynamic microfluidic trapping are discussed. Based on their combination, on‐chip applications beyond SERS are presented, and insight can be gained into the commercialization of portable SERS chips.