A Fully Integrated Miniaturized Optical Biosensor for Fast and Multiplexing Plasmonic Detection of High- and Low-Molecular-Weight Analytes.

Optical biosensors based on plasmonic sensing schemes combine high sensitivity and selectivity with label-free detection. However, the use of bulky optical components is still hampering the possibility of obtaining miniaturized systems required for analysis in real settings. Here, a fully miniaturized optical biosensor prototype based on plasmonic detection is demonstrated, which enables fast and multiplex sensing of analytes with high- and low molecular weight (80 000 and 582 Da) as quality and safety parameters for milk: a protein (lactoferrin) and an antibiotic (streptomycin). The optical sensor is based on the smart integration of: i) miniaturized organic optoelectronic devices used as light-emitting and light-sensing elements and ii) a functionalized nanostructured plasmonic grating for highly sensitive and specific localized surface plasmon resonance (SPR) detection. The sensor provides quantitative and linear response reaching a limit of detection of 10-4 refractive index units once it is calibrated by standard solutions. Analyte-specific and rapid (15 min long) immunoassay-based detection is demonstrated for both targets. By using a custom algorithm based on principal-component analysis, a linear dose-response curve is constructed which correlates with a limit of detection (LOD) as low as 3.7 µg mL-1 for lactoferrin, thus assessing that the miniaturized optical biosensor is well-aligned with the chosen reference benchtop SPR method.

Refractive Index Dependence of Fluorescence Enhancement in a Nanostructured Plasmonic Grating.

Plasmonic gratings are attracting huge interest in the context of realizing sensors based on surface-enhanced fluorescence. The grating features control the plasmonic modes and consequently have a strong effect on the fluorescence response. Within this framework, we focused on the use of a buffer solution flowing across the grating active surface to mimic a real measurement. The refractive index of the surrounding medium is therefore altered, with a consequent modification of the resonance conditions. The result is a shift in the spectral features of the fluorescence emission accompanied by a reshaping of the fluorescence emission in terms of spectral weight and direction.

Plasmonic Modes and Fluorescence Enhancement Coupling Mechanism: A Case with a Nanostructured Grating.

The recent development and technological improvement in dealing with plasmonic metasurfaces has triggered a series of interesting applications related to sensing challenges. Fluorescence has been one of the most studied tools within such a context. With this in mind, we used some well characterized structures supporting plasmonic resonances to study their influence on the emission efficiency of a fluorophore. An extended optical analysis and a complementary investigation through finite-difference time-domain (FDTD) simulations have been combined to understand the coupling mechanism between the excitation of plasmonic modes and the fluorescence absorption and emission processes. The results provide evidence of the spectral shape dependence of fluorescence on the plasmonic field distribution together with a further relationship connected with the enhancement of its signal. It has made evident that the spectral region characterized by the largest relative enhancement closely corresponds to the strongest signatures of the plasmonic modes, as described by both the optical measurements and the FDTD findings.

Plasma Fabrication and SERS Functionality of Gold Crowned Silicon Submicrometer Pillars.

Sequential plasma processes combined with specific lithographic methods allow for the fabrication of advanced material structures. In the present work, we used self-assembled colloidal monolayers as lithographic structures for the conformation of ordered Si submicrometer pillars by reactive ion etching. We explored different discharge conditions to optimize the Si pillar geometry. Selected structures were further decorated with gold by conventional sputtering, prior to colloidal monolayer lift-off. The resulting structures consist of a gold crown, that is, a cylindrical coating on the edge of the Si pillar and a cavity on top. We analysed the Au structures in terms of electronic properties by using X-ray absorption spectroscopy (XAS) prior to and after post-processing with thermal annealing at 300 °C and/or interaction with a gold etchant solution (KI). The angular dependent analysis of the plasmonic properties was studied with Fourier transformed UV-vis measurements. Certain conditions were selected to perform a surface enhanced Raman spectroscopy (SERS) evaluation of these platforms with two model dyes, prior to confirming the potential interest for a well-resolved analysis of filtered blood plasma.

Porous Silicon Bragg Reflector and 2D Gold-Polymer Nanograting: A Route Towards a Hybrid Optoplasmonic Platform.

Photonic and plasmonic systems have been intensively studied as an effective means to modify and enhance the electromagnetic field. In recent years hybrid plasmonic-photonic systems have been investigated as a promising solution for enhancing light-matter interaction. In the present work we present a hybrid structure obtained by growing a plasmonic 2D nanograting on top of a porous silicon distributed Bragg reflector. Particular attention has been devoted to the morphological characterization of these systems. Electron microscopy images allowed us to determine the geometrical parameters of the structure. The matching of the optical response of both components has been studied. Results indicate an interaction between the plasmonic and the photonic parts of the system, which results in a localization of the electric field profile.

Biofouling Properties of Nitroxide-Modified Amorphous Carbon Surfaces.

Amorphous carbon films exhibit attractive optical and surface properties. In this work, modified amorphous carbon films incorporating nitroxide groups (α-CNO) have been obtained by searching for a condensed analogue to classical soft antifouling materials. Thin films deposited by reactive magnetron sputtering in air discharges at varying power conditions were characterized by ellipsometry, atomic force microscopy, and water contact angle. Plasma power was observed to activate the densification and roughness of nanograined films. Most hydrophilic films deposited at 30 W exhibited the lowest refractive index, negligible optical absorption in the vis-IR, and presented a close to stoichiometric C2NO composition, as derived from X-ray photoelectron spectroscopy. Micropatterns prepared by photolithography validated the transparency-hydrophilicity of the α-CNO, as observed by water condensation contrast imaging. An albumin adsorption experiment evaluated through fluorescence revealed that α-CNO behaves as antifouling with respect to Si. Such thin antifouling films are of interest for the initiation of immobilization cascades in imaging surface plasmon resonance, where they have confirmed their antifouling contrast enhancement role. These results illustrate that the combination of a nanorough surface with nitroxide chemistry induces an antifouling behavior. In association with the optical transparency, the results invite the exploration of the bioengineering dimension of α-CNO films.

Synergic combination of the sol-gel method with dip coating for plasmonic devices.

Biosensing technologies based on plasmonic nanostructures have recently attracted significant attention due to their small dimensions, low-cost and high sensitivity but are often limited in terms of affinity, selectivity and stability. Consequently, several methods have been employed to functionalize plasmonic surfaces used for detection in order to increase their stability. Herein, a plasmonic surface was modified through a controlled, silica platform, which enables the improvement of the plasmonic-based sensor functionality. The key processing parameters that allow for the fine-tuning of the silica layer thickness on the plasmonic structure were studied. Control of the silica coating thickness was achieved through a combined approach involving sol-gel and dip-coating techniques. The silica films were characterized using spectroscopic ellipsometry, contact angle measurements, atomic force microscopy and dispersive spectroscopy. The effect of the use of silica layers on the optical properties of the plasmonic structures was evaluated. The obtained results show that the silica coating enables surface protection of the plasmonic structures, preserving their stability for an extended time and inducing a suitable reduction of the regeneration time of the chip.

Polymer nanopillar-gold arrays as surface-enhanced Raman spectroscopy substrate for the simultaneous detection of multiple genes.

In our study, 2D nanopillar arrays with plasmonic crystal properties are optimized for surface-enhanced Raman spectroscopy (SERS) application and tested in a biochemical assay for the simultaneous detection of multiple genetic leukemia biomarkers. The special fabrication process combining soft lithography and plasma deposition techniques allows tailoring of the structural and chemical parameters of the crystal surfaces. In this way, it has been possible to tune the plasmonic resonance spectral position close to the excitation wavelength of the monochromatic laser light source in order to maximize the enhancing properties of the substrate. Samples are characterized by scanning electron microscopy and reflectance measurements and tested for SERS activity using malachite green. Besides, as the developed substrate had been prepared on a simple glass slide, SERS detection from the support side is also demonstrated. The optimized substrate is functionalized with thiol-modified capture oligonucleotides, and concentration-dependent signal of the target nucleotide is detected in a sandwich assay with labeled gold nanoparticles. Gold nanoparticles functionalized with different DNA and various Raman reporters are applied in a microarray-based assay recognizing a disease biomarker (Wilms tumor gene) and housekeeping gene expressions in the same time on spatially separated microspots. The multiplexing performance of the SERS-based bioassay is illustrated by distinguishing Raman dyes based on their complex spectral fingerprints.

Multiplexed label-free optical biosensor for medical diagnostics.

This paper describes a new multiplexed label-free biosensor. The detection technology is based on nanostructured gold-polymer surfaces. These surfaces support surface plasmon resonance modes that can be probed by a miniaturized optical setup. The optical characterization of the sensing chip shows the sensitivity and the limit-of-detection to refractive index changes. Moreover, by studying the progressive adhesion of molecular monolayers of polyelectrolytes, the decay of the plasmonic mode electric field above the surface has been reconstructed. A multiplexed label-free biosensing device is then described and characterized in terms of sensitivity, lateral resolution, and sensitivity to a model biological assay. The sensitivity in imaging mode of the device is of the order of 10-6 refractive index units, while the measured lateral resolution is 6.25 µm within a field of view of several tenths of mm2, making the instrument unique in terms of multiplexing capability. Finally, the proof-of-concept application of the technology as a point-of-care diagnostic tool for an inflammatory marker is demonstrated.

Optical Sensitivity Gain in Silica-Coated Plasmonic Nanostructures.

Ultrathin films of silica realized by sol-gel synthesis and dip-coating techniques were successfully applied to predefined metal/polymer plasmonic nanostructures to spectrally tune their resonance modes and to increase their sensitivity to local refractive index changes. Plasmon resonance spectral shifts up to 100 nm with slope efficiencies of ∼8 nm/nm for increasing layer thickness were attained. In the ultrathin layer regime (<10 nm), which could be reached by suitable dilution of the silica precursors and optimization of the deposition speed, the sensitivity of the main plasmonic resonance to refractive index changes in aqueous solution could be increased by over 50% with respect to the bare plasmonic chip. Numerical simulations supported experimental data and unveiled the mechanism responsible for the optical sensitivity gain, proving an effective tool in the design of high-performance plasmonic sensors.