Analyte Co-localization at Electromagnetic Gap Hot-Spots for Highly Sensitive (Bio)molecular Detection by Plasmon Enhanced Spectroscopies

Electromagnetic hot-spots at ultranarrow plasmonic nanogaps carry immense potential to drive detection limits down to few molecules in sensors based on surface-enhanced Raman or fluorescence spectroscopies. However, leveraging the EM hot-spots requires access to the gaps, which in turn depends on the size of the analyte in relation to gap distances. Herein, we leverage a well-calibrated process based on self-assembly of block copolymer colloids on a full-wafer level to produce high-density plasmonic nanopillar arrays exhibiting a large number (>10¹⁰ cm^–2) of uniform interpillar EM hot-spots. The approach allows convenient handles to systematically vary the interpillar gap distances down to a sub-10 nm regime. The results show compelling trends of the impact of analyte dimensions in relation to the gap distances toward their leverage over interpillar hot-spots and the resulting sensitivity in SERS-based molecular assays. Comparing the detection of labeled proteins in surface-enhanced Raman and metal-enhanced fluorescence configurations further reveal the relative advantage of fluorescence over Raman detection while encountering the spatial limitations imposed by the gaps. Quantitative assays with limits of detection down to picomolar concentrations are realized for both small organic molecules and proteins. The well-defined geometries delivered by a nanofabrication approach are critical to arriving at realistic geometric models to establish meaningful correlation between the structure, optical properties, and sensitivity of nanopillar arrays in plasmonic assays. The findings emphasize the need for the rational design of EM hot-spots that takes into account the analyte dimensions to drive ultrahigh sensitivity in plasmon-enhanced spectroscopies.