96-Well Plasmonic Sensing with Nanohole Arrays

Abstract

A multiwell plasmonic reader was designed and validated for higher throughput analysis of biological interactions with a platform of the same size as standard 96-well plates. While the plasmonic sensor can be read with standard 96-well plate readers, a custom 96-well plate reader was designed to analyze nanohole arrays at high incident angles required for higher sensitivity. Gold nanohole arrays were manufactured on a 4 in. glass wafer using a photolithographic process. In comparison to single channel measurements with nanohole arrays fabricated with nanosphere lithography, the nanohole array sensors greatly enhanced the signal-to-noise ratio of the plasmonic signal and precision of the measurements with the multiwell plate system. As proof of concept, the detection of IgG in the low nanomolar range was achieved with the multiwell plate reader. The multiwell plasmonic plate reader was also applied to the screening of several prostate specific (PSA) antibodies for secondary detection of PSA and for the analysis of an anticancer drug through a competitive assay between methotrexate (MTX) and folic acid Au nanoparticle (FaNP) for human dihydrofolate reductase (hDHFR). The multiwell plasmonic reader based on nanohole array technology offers the rapid, versatile, sensitive, and simple high throughput detection of biomolecules.

High throughput LSPR and SERS analysis of aminoglycoside antibiotics

“…30 The main difference with the setup described here is the use of plain glass coverslips as opposed to nanohole array surfaces, to create a simple multi-well setup for solution based sensing. Varying concentrations of tobramycin was added to 20 nm Au-DTNB in a 1:1 ratio to a final volume of 100 μL.…”

Quantitative Comparison of Protein Adsorption and Conformational Changes on Dielectric-Coated Nanoplasmonic Sensing Arrays

“…Within this scope, proteins have become a popular subject of inquiry as protein adsorption onto a solid support is often accompanied by substrate-induced denaturation and resulting conformational changes. Indeed, while most nanoplasmonic sensing works involving proteins have focused on end-point detection [ 19 , 20 ], there is a growing number of studies that have exploited the high surface sensitivity of LSPR measurement strategies to investigate protein-protein interactions [ 21 , 22 , 23 , 24 , 25 ], protein interactions with small molecules [ 26 , 27 , 28 , 29 ], sugars [ 30 , 31 , 32 ], and drugs [ 33 , 34 ], and protein conformational changes triggered by an environmental stimuli [ 9 ]. In recent years, the utility of LSPR sensors to study protein adsorption has become greatly expanded through the introduction of indirect nanoplasmonic sensing (INPS), which involves coating plasmonic metal nanoparticles with a dielectric material of interest [ 35 ].…”

Plasmon properties and hybridization effects in silicene

“…Recent studies have thoroughly characterized the plasmon dispersions * antonello.sindona@fis.unical.it in MG [17][18][19][20][21][22] and some other graphene-related materials, such as bilayer graphene (BLG) [23,24] and graphene nanoribbons (GNRs) arranged in regular planar arrays,which, unlike MG, offer geometrically controllable band gaps [25][26][27]. Further advances are expected from the analysis of nanocarbon-metal interfaces together with associated application developments in biological sensing, optical signal processing and quantum information technology [3,[28][29][30][31][32].…”