“…In particular, nanoscale junctions between metal nanoparticles, known as “hotspots”, can dramatically enhance local electromagnetic (EM) fields and thus allow ultrasensitive analysis of molecules located inside the hotspots . However, owing to the near-field nature of SERS phenomenon, the enhancement factors (EFs) in the hotspots are highly sensitive to the size, shape, and spacing of the nanoparticles, − as well as local dielectric environments. − As a result, quantitative SERS analysis has been a major challenge in practical applications because of the difficulty in obtaining uniform and reproducible SERS substrates. , Recently, self-assembled nanoparticle monolayers with high-density and uniform hotspots have attracted great interests as SERS substrates. ,− In particular, self-assembled gold nanoparticle (AuNP) monolayers offer the advantages of high plasmonic activity in the visible to near-infrared regions, chemical stability, well-tunable nanostructures, , and simplicity and scalability of their preparation. − However, the EFs of self-assembled AuNP monolayers can still vary by orders of magnitude due to sample and instrument variations, such as laser power, integration time, and polarization of incident light …”