Fluorescence signal amplification by optical reflection in metal-coated nanowells

A significant amplification in the fluorescence signal is demonstrated when measured in metal (aluminum)-coated fluidic wells with volumes on the order of a nanoliter or smaller (nanowells). Photolithographic and wet etching procedures were used to fabricate these nanowells on glass substrates followed by vapor deposition of an aluminum layer on them. The fluorescence signal recorded in these structures was enhanced due to the reflection of the incident and emitted radiation by the metal layer as well as focusing of this light by the curvature of the well surface. While the first effect amplified the background signal in the entire assay chamber, the latter one produced signal hotspots around the edges and center of the nanowell. In this work, we were able to realize over a 20-fold enhancement in the fluorescence signal upon quantitating it at the central hotspot of an aluminum-coated circular nanowell with a depth and photo-patterned diameter of 30 µm and 38 µm, respectively. More interestingly, our experiments indicate that this enhancement factor may be further improved by optimizing the curvature of the nanowell surface to merge all the signal hotspots within a smaller detection zone. Finally, quantitative assays using horseradish peroxidase samples were performed on the reported signal enhancement platform to further demonstrate its utility for making sensitive analytical measurements.