Nanobiosensors for in vitro and in vivo analysis of biomolecules.

This chapter presents as a proof of concept the development of a nanosensor based on the localized surface plasmon resonance for the analysis of biomolecules. The method presented take advantage of the plasmon generated in the surrounding of gold nanoparticles (i.e., 100 nm) for the specific interaction between antigen and antibody. The procedure for the optimization of an assay for the determination of biomolecules consisted mainly of four steps. First, the immobilization of gold nanoparticles over the glass surface using the appropriate ratio, concentration and time-contact of amino-sylilating agent, and nonreactive sylilating agent. Next, the suitable concentration of coating antigen in order to obtain the maximum signal LSPR. Following this step, the interaction between antigen and antibody (specific antibody) is evaluated by measuring the signal LSPR. Finally, a calibration curve was obtained for the detection of a small organic molecule such as stanozolol using this nanobiosensor. As a proof of concept, the use of a model is performed that in this case is for the detection of an anabolic androgenic steroid, such as stanozolol which is banned for the European Commission (EC) as a growth promoter and for the World Anti-Doping Agency (WADA) as a doping agent. The nanosensor developed demonstrates its feasibility for screening purposes due to the limit of detection achieved (0.7 µg/L) is under the MRPL required for both organizations (10 µg/L). A protocol such as that presented here may be generally applied for the analysis of other pollutant such as pesticides or antibiotics, or for biomedical applications for the analysis of biomarkers using the LSPR principle using gold nanoparticles (i.e., 30-120 nm).

Embedded optical micro/nano-fibers for stable devices.

We present a technique to embed silica micro and nanofibers in low-index material (Teflon) using an inexpensive and straightforward fabrication process based on spin coating. The optical properties of the silica micro/nano-fibers have been investigated when they are bare or completely or partially embedded. Optical degradation occurs in bare fibers with diameters smaller than twice the wavelength of the guided light, thus making protection through embedding necessary. Our results also show that completely embedded fibers do not degrade over a long time, while partially embedded fibers can preserve the large evanescent waves without undergoing considerable degradation, which would be further reduced or even become negligible with functional overlayers. The results represent a step forward toward the development of durable and stable devices based on optical micro/nano fibers.

Refractometry based on a photonic crystal fiber interferometer.

We report a simple and compact modal interferometer for applications in refractometry. The device consists of a stub of large-mode-area photonic crystal fiber (PCF) spliced between standard single-mode fibers. In the splice regions the voids of the PCF are fully collapsed, thus allowing the coupling and recombination of PCF core and cladding modes. The device is highly stable over time, has low temperature sensitivity, and is suitable for measuring indices in the 1.330-1.440 range. The measure of the refractive index is carried out by monitoring the shift of the interference pattern.

Photonic crystal fiber interferometer for chemical vapor detection with high sensitivity.

We report an in-reflection photonic crystal fiber (PCF) interferometer which exhibits high sensitivity to different volatile organic compounds (VOCs), without the need of any permeable material. The interferometer is compact, robust, and consists of a stub of PCF spliced to standard optical fiber. In the splice the voids of the PCF are fully collapsed, thus allowing the excitation and recombination of two core modes. The device reflection spectrum exhibits sinusoidal interference pattern which shifts differently when the voids of the PCF are infiltrated with VOC molecules. The volume of voids responsible for the shift is less than 600 picoliters whereas the detectable levels are in the nanomole range.

Localized surface plasmon resonance effects on the magneto-optical activity of continuous Au/Co/Au trilayers.

We study how the magneto-optical activity in polar configuration of continuous Au/Co/Au trilayers is affected by the excitation of localized plasmon resonances of an array of Au nanodiscs fabricated on top of them over a dielectric SiO(2) spacer. We show that the effect of the nanodiscs array is twofold. First, it optimizes the absorption of light at specific photon energies corresponding to the localized surface plasmon excitation of the array, modifying the reflectivity of the system (we define this effect as the purely optical contribution). Second, upon localized plasmon resonance excitation, the electromagnetic field in the whole system is redistributed, and an enhanced magneto-optical activity occurs at those energies where the electromagnetic field in the magnetic layer is increased (this effect is identified as the purely magneto-optical contribution of the nanodiscs array).

Colloidal-based localized surface plasmon resonance (LSPR) biosensor for the quantitative determination of stanozolol.

This work reports the systematic preparation of biosensors through the use of functionalized glass substrates, noble metal gold colloid, and measurement by localized surface plasmon resonance (LSPR). Glass substrate was modified through chemical silanization, and the density of gold colloid was carefully controlled by optimizing the conditions of silanization through the use of mixed silanes and selective mixing procedures. At this point, samples were exposed to bioreagents and changes in the shallow dielectric constant around the particles were observed by dark-field spectroscopy. Biological binding of high affinity systems (biotin/streptavidin and antigen/antibody) was subsequently investigated by optimizing coating layers, receptor concentration profiling, and finally quantitative determination of the analyte of interest, which in this case was a small organic molecule-the widely used, synthetic anabolic steroid called stanozolol. For this system, high specificity was achieved (>97%) through extensive nonspecific binding tests, with a sensitivity measurable to a level below the minimum required performance level (MRPL) as determined by standard chromatographic methods. Analytical best-fit parameters of Hillslope and regression coefficient are also commented on for the final LSPR biosensor. The LSPR biosensor showed good reproducibility (<5% RSD) and allowed for rapid preparation of calibration curves and determination of the analyte (measurement time of each sample ca. 2 min). As an alternative method for quantitative steroidal analysis, this approach significantly simplifies the detection setup while reducing the cost of analysis. In addition the system maintains comparable sensitivity to standard surface plasmon resonance methods and offers great potential for miniaturization and development of multiplexed devices.

A hybrid light source with integrated inorganic light-emitting diode and organic polymer distributed feedback grating.

We report a compact light source that incorporates a semiconductor light-emitting diode, nanostructured distributed feedback (DFB) Bragg grating and spin-coated thin conjugated polymer film. With this hybrid structure, we transferred electrically generated 390 nm ultraviolet light to an organic polymer via optical pumping and out-couple green luminescence to air through a second-order DFB grating. We demonstrate the feasibility of electrically driven, hybrid, compact light-emitting devices and lasers in the visible range.

Simple all-microstructured-optical-fiber interferometer built via fusion splicing.

We report a compact and stable all-microstructured-optical-fiber interferometer built with two fusion splices separated a few centimeters from each other. The air-holes of the fiber are intentionally collapsed in the vicinity of the splices. This broadens the propagating optical mode, allowing coupling of two modes in the section between the splices. A truly sinusoidal interference pattern was observed from 800 nm to 1600 nm with fringe visibility reaching 80%. The fringe spacing was inversely proportional to the distance between the splices. The potential of the device for sensing applications is demonstrated.

Tunable optical sorting and manipulation of nanoparticles via plasmon excitation.

We numerically investigate the optical forces exerted by an incident light beam on Rayleigh metallic particles over a dielectric substrate. In analogy with atom manipulation, we identify two different trapping regimes depending on whether the illumination is performed within the plasmon band or out of it. By adjusting the incident wavelength, the particles can be selectively guided, or immobilized, at the substrate interface.

Surface plasmon radiation forces.

We report the first experimental observation of momentum transfer from a surface plasmon to a single dielectric sphere. Using a photonic force microscope, we measure the plasmon radiation forces on different polystyrene beads as a function of their distance from the metal surface. We show that the force magnitude at resonance is strongly enhanced compared to a nonresonant illumination. Measurements performed as a function of the probe particle size indicate that optical manipulation by plasmon fields has a strong potential for optical sorting.

Quantitative detection of doping substances by a localised surface plasmon sensor.

Within this communication, consistent evidence of a quantitative biosensing principle for steroidal residue analysis is presented. Our approach uses a simple method for the quantitative determination of an anabolic agent called stanozolol (Sz). Sz (Mw 328) is widely used in sports, horse racing and as a growth promoter in animals for human consumption. Through the use of localised surface plasmons (LSPs), sustained by three-dimensional noble metal nanostructures, we have developed a highly specific, label-less immunosensor for the detection of this small organic molecule to low levels (nM range). A main practical advantage over conventional flat extended film surface plasmon resonance (SPR) systems is the simplicity of the optical configuration, since there is no need for cumbersome total internal reflection illumination, thus making integration easier. In addition, the active area of the LSP-based sensor is smaller, decreasing the minimum detectable number of molecules involved in the binding event. Assay times are short and the set-up is comprised of relatively cheap instrumentation. Detection levels found here are comparable with SPR, even at this early stage of development and with further modifications, we envisage sensing down to pM (10(-12)) levels.

Microstructured optical fiber coated with thin films for gas and chemical sensing.

We propose the use of tapered microstructured fibers with collapsed air-holes coated with thin layers for gas sensing. The collapsing of the holes allows having access to the evanescent fields which can be absorbed or attenuated with gas-permeable thin films. On the other hand, a section of the holey fiber is transformed into a solid multimode fiber. The beating between the multiple modes of the latter makes the transmission spectra of the device to exhibit an oscillatory pattern. This evanescent-fields-plus-modal-interferometer structure may offer interesting properties for gas and chemical sensing. As an example we demonstrate a hydrogen sensor.

Cumulative plasmon field enhancement in finite metal particle chains.

The dramatic field enhancement at the extremity of finite chains of strongly coupled gold nanoparticles illuminated under total internal reflection is investigated numerically. We demonstrate that high enhancement factors can be achieved by exploiting the in-plane forward scattering of the particles, with geometries achievable by state-of-the-art lithographic techniques.

Radiation forces on a Rayleigh dielectric sphere in a patterned optical near field.

We report on the study of the radiation forces exerted on a Rayleigh dielectric particle by a patterned optical near-field landscape at an interface decorated with resonant gold nanostructures. This configuration allows for the generation of a large array of surface subwavelength optical traps from an extended collimated beam, which may be of interest for parallel optical manipulation and sorting of submicrometer objects.

Electromagnetic coupling between a metal nanoparticle grating and a metallic surface.

The electromagnetic coupling between a two-dimensional grating of resonant gold nanoparticles and a gold metallic film is investigated. We report on the observation of multipeaks in the extinction spectra attributed to resonant modes of the hybrid system, resulting from the coupling between the localized plasmon of the nanoparticles with the underlying surface plasmon mode. Simulations based on the Fourier modal method give good agreement with the experimental measurements and allow for the identification of the respective contributions.

Sub-wavelength patterning of the optical near-field.

We report the sub-wavelength patterning of the optical near-field by total internal reflection illumination of a regular array of resonant gold nano-particles. Under appropriate conditions, the in-plane coupling between Localized Surface Plasmon (LSP) fields gives rise to sub-wavelength light spots between the structures. Measurements performed with an Apertureless Scanning Near-Field Optical Microscope (ASNOM) show a good agreement with theoretical predictions based on the Green dyadic method. This concept might offer a convenient way to elaborate extended optical trap landscapes for manipulation of sub-micrometer systems.

Optical sensing based on plasmon coupling in nanoparticle arrays.

The performance of bi-periodic arrays of gold nano-particles for molecular sensing applications is studied using the Fourier Modal Method (FMM). We show that the electromagnetic coupling between the particles can be optimized to increase their sensitivity to a weak change of the shallow dielectric environment. Especially, arrays whose elementary cell consists of a dimer of two closely packed particles are found to be at least three times more sensitive than single particle arrays.