Insights into Photopolymerization at the Nanoscale Using Surface Plasmon Resonance Imaging.

Near-field photopolymerization (NFPP) driven by surface plasmon resonance has attracted increasing attention in nanofabrication. This interest comes from the nanometer-scale control of polymer thickness, due to the confinement of the evanescent wave within a highly restricted volume at the surface. In this study, a novel approach using a multi-spectral surface plasmon resonance instrument is presented that gives access to real-time images of polymer growth during NFPP with nanometer sensitivity. Using the plasmonic evanescent wave for both polymerization and real-time sensing, the influence of irradiance, concentration of dye, and initiator are investigated on the threshold energy and kinetics of NFPP. How oxygen inhibition in the near field strongly affects photopolymerization is highlighted, more than in the far field.

Performance improvement of plasmonic sensors using a combination of AC electrokinetic effects for (bio)target capture.

Analytes concentration techniques are being developed with the appealing expectation to boost the performance of biosensors. One promising method lies in the use of electrokinetic forces. We present hereafter a new design for a microstructured plasmonic sensor which is obtained by conventional microfabrication techniques, and which can easily be adapted on a classical surface plasmon resonance imaging (SPRI) system without further significant modification. Dielectrophoretic trapping and electro-osmotic displacement of the targets in the scanned fluid are performed through interdigitated 200 µm wide gold electrodes that also act as the SPR-sensing substrate. We demonstrate the efficiency of our device's collection capabilities for objects of different sizes (200 nm and 1 µm PS beads, as well as 5-10 µm yeast cells). SPRI is relevant for the spatial analysis of the mass accumulation at the electrode surface. We demonstrate that our device overcomes the diffusion limit encountered in classical SPR sensors thanks to rapid collection capabilities (<1 min) and we show a consequent improvement of the detection limit, by a factor >300. This study of an original device combining SPRI and electrokinetic forces paves the way to the development of fully integrated active plasmonic sensors with direct applications in life sciences, electrochemistry, environmental monitoring and agri-food industry.

Spatial resolution versus contrast trade-off enhancement in high-resolution surface plasmon resonance imaging (SPRI) by metal surface nanostructure design.

Surface plasmon resonance imaging (SPRI) is an optical near-field method used for mapping the spatial distribution of chemical/physical perturbations above a metal surface without exogenous labeling. Currently, the majority of SPRI systems are used in microarray biosensing, requiring only modest spatial resolution. There is increasing interest in applying SPRI for label-free near-field imaging of biological cells to study cell/surface interactions. However, the required resolution (sub-µm) greatly exceeds what current systems can deliver. Indeed, the attenuation length of surface plasmon polaritons (SPP) severely limits resolution along one axis, typically to tens of µm. Strategies to date for improving spatial resolution result in a commensurate deterioration in other imaging parameters. Unlike the smooth metal surfaces used in SPRI that support purely propagating surface modes, nanostructured metal surfaces support "hybrid" SPP modes that share attributes from both propagating and localized modes. We show that these hybrid modes are especially well-suited to high-resolution imaging and demonstrate how the nanostructure geometry can be designed to achieve sub-µm resolution while mitigating the imaging parameter trade-off according to an application-specific optimum.

Fisheye-Based Method for GPS Localization Improvement in Unknown Semi-Obstructed Areas.

A precise GNSS (Global Navigation Satellite System) localization is vital for autonomous road vehicles, especially in cluttered or urban environments where satellites are occluded, preventing accurate positioning. We propose to fuse GPS (Global Positioning System) data with fisheye stereovision to face this problem independently to additional data, possibly outdated, unavailable, and needing correlation with reality. Our stereoscope is sky-facing with 360° × 180° fisheye cameras to observe surrounding obstacles. We propose a 3D modelling and plane extraction through following steps: stereoscope self-calibration for decalibration robustness, stereo matching considering neighbours epipolar curves to compute 3D, and robust plane fitting based on generated cartography and Hough transform. We use these 3D data with GPS raw data to estimate NLOS (Non Line Of Sight) reflected signals pseudorange delay. We exploit extracted planes to build a visibility mask for NLOS detection. A simplified 3D canyon model allows to compute reflections pseudorange delays. In the end, GPS positioning is computed considering corrected pseudoranges. With experimentations on real fixed scenes, we show generated 3D models reaching metric accuracy and improvement of horizontal GPS positioning accuracy by more than 50%. The proposed procedure is effective, and the proposed NLOS detection outperforms CN0-based methods (Carrier-to-receiver Noise density).

Directional surface enhanced Raman scattering on gold nano-gratings.

Directional plasmon excitation and surface enhanced Raman scattering (SERS) emission were demonstrated for 1D and 2D gold nanostructure arrays deposited on a flat gold layer. The extinction spectrum of both arrays exhibits intense resonance bands that are redshifted when the incident angle is increased. Systematic extinction analysis of different grating periods revealed that this band can be assigned to a propagated surface plasmon of the flat gold surface that fulfills the Bragg condition of the arrays (Bragg mode). Directional SERS measurements demonstrated that the SERS intensity can be improved by one order of magnitude when the Bragg mode positions are matched with either the excitation or the Raman wavelengths. Hybridized numerical calculations with the finite element method and Fourier modal method also proved the presence of the Bragg mode plasmon and illustrated that the enhanced electric field of the Bragg mode is particularly localized on the nanostructures regardless of their size.

Generalized analytical model based on harmonic coupling for hybrid plasmonic modes: comparison with numerical and experimental results.

Metal nanoparticle arrays have proved useful for different applications due to their ability to enhance electromagnetic fields within a few tens of nanometers. This field enhancement results from the excitation of various plasmonic modes at certain resonance frequencies. In this article, we have studied an array of metallic nanocylinders placed on a thin metallic film. A simple analytical model is proposed to explain the existence of the different types of modes that can be excited in such a structure. Owing to the cylinder array, the structure can support localized surface plasmon (LSP) modes. The LSP mode couples to the propagating surface plasmon (PSP) mode of the thin film to give rise to the hybrid lattice plasmon (HLP) mode and anti-crossing phenomenon. Due to the periodicity of the array, the Bragg modes (BM) are also excited in the structure. We have calculated analytically the resonance frequencies of the BM, LSP and the corresponding HLP, and have verified the calculations by rigorous numerical methods. Experimental results obtained in the Kretschmann configuration also validate the proposed analytical model. The dependency of the resonance frequencies of these modes on the structural parameters such as cylinder diameter, height and the periodicity of the array is shown. Such a detailed study can offer insights on the physical phenomenon that governs the excitation of various plasmonic modes in the system. It is also useful to optimize the structure as per required for the different applications, where such types of structures are used.

Microrheology and structural quantification of hypercoagulable clots.

Hypercoagulability is a pathology that remains difficult to explain today in most cases. It is likely due to a modification of the conditions of polymerization of the fibrin, the main clot component. Using passive microrheology, we measured the mechanical properties of clots and correlated them under the same conditions with structural information obtained with confocal microscopy. We tested our approach with known alterations: an excess of fibrinogen and of coagulation Factor VIII. We observed simultaneously a rigidification and densification of the fibrin network, showing the potential of microrheology for hypercoagulability diagnosis.

Biochip data normalization using multifunctional probes.

Using a biochip with stable probe functionalization and a detection system capable of real time measurements, it is demonstrated that acquired probe-target interaction data are more reproducible in time--on a given probe spot using sequential target runs--than in space, using many probe spot replicates on the biochip in one single parallel target run. To increase the biochip data precision, a normalization method that quantifies and corrects the surface inhomogeneity without the use of complex data post-processing has been developed. This simple and effective method is based on adding a common reactive group to all probes and quantifying the biochip response to a calibration target, thus quantifying the spatial heterogeneity in the biosensor responsiveness. The usefulness of such methodology, which can be easily generalized, is demonstrated in the model case of DNA:DNA interactions, using a surface plasmon resonance imaging system as the dynamical reader. The biochips are based on streptavidin biochemically functionalized gold films onto which biotinylated ssDNA probe sequences, related to cystic fibrosis genotyping, are spotted. This normalization method provides high gain in data precision and allows, in this example, unambiguous genotyping of SNP, including discrimination of the heterozygote case from the two homozygote cases.

Surface micropatterning for the formation of an in vitro functional endothelial model for cell-based biosensors.

Label-free biosensing, such as with surface plasmon resonance (SPR), is a highly efficient method for monitoring the responses of living cells exposed to pharmacological agents and biochemical stimuli in vitro. Conventional cell culture protocols used in cell-based biosensing generally provide little direct control over cell morphologies and phenotypes. Surface micropatterning techniques have been exploited for the controlled immobilization and establishment of well-defined cell morphologies and phenotypes. In this article, surface adhesion micropatterns are used to control the adhesion of endothelial cells within adjacent hexagonal microstructures to promote the emergence of a well-controlled and standardized cell layer phenotype onto SPR sensor surfaces. We show that the formation of cell-cell junctions can be controlled by tuning the inter-cellular spacing in groups of 3 neighbouring cells. Fluorescence microscopy was used to confirm the formation of vascular endothelium cadherin junctions, a structural marker of a functional endothelium. In order to confirm the functionality of the proposed model, the response to thrombin, a modulator of endothelium integrity, was monitored by surface plasmon resonance imaging (SPRI). Experiments demonstrate the potential of the proposed model as a primary biological signal transducer for SPRI-based analysis, with potential applications in cell biology, pharmacology and diagnostic.

Spatially-Localized Functionalization on Nanostructured Surfaces for Enhanced Plasmonic Sensing Efficacy.

This work demonstrates the enhancement in plasmonic sensing efficacy resulting from spatially-localized functionalization on nanostructured surfaces, whereby probe molecules are concentrated in areas of high field concentration. Comparison between SERS measurements on nanostructured surfaces (arrays of nanodisks 110 and 220 nm in diameter) with homogeneous and spatially-localized functionalization with thiophenol demonstrates that the Raman signal originates mainly from areas with high field concentration. TERS measurements with 10 nm spatial resolution confirm the field distribution profiles predicted by the numerical modeling. Though this enhancement in plasmonic sensing efficacy is demonstrated with SERS, results apply equally well to any type of optical/plasmonic sensing on functionalized surfaces with nanostructuring.

Angulo-spectral surface plasmon resonance imaging of nanofabricated grating surfaces.

We present a surface plasmon resonance imaging (SPRI) setup, based on the Kretschmann configuration, capable of simultaneously acquiring the complete spectral and angular plasmonic reflectivity response on all points of the sensing area. Several line poly(methyl methacrylate) grating regions were fabricated on a thin-film gold surface and characterized with this SPRI system. Reflectivity maps of the corrugated regions showing plasmon bandgaps were obtained to illustrate the capability of the setup.

Motion artifact suppression in full-field optical coherence tomography.

Significant motion artifacts limit the performance of conventional full-field optical coherence tomography (FF-OCT) for in-vivo imaging. We present a theoretical and experimental study of those limitations. A new FF-OCT system suppressing most of artifacts due to sample motions is demonstrated using instantaneous phase shifting with nonpolarizing optics and pulsed illumination. The experimental setup is based on a Linnik-type interferometer illuminated by the superluminescence emission from a Ti:Al(2)O(3) waveguide crystal. En face tomographic images are calculated as a combination of two phase-opposed interferometric images acquired simultaneously by two CCD cameras placed at both outputs of the interferometer, with a spatial resolution of 0.8 microm x 1.6 microm (axial x transverse) and a detection sensitivity of approximately 60 dB.

Polarimetric surface plasmon resonance imaging biosensor.

We report the realization of a polarimetric surface plasmon resonance imaging system capable of dynamically resolving a change in the optical anisotropy of biochemical films. Anisotropies as small as 10(-3) refractive index unit on nanometer-thick samples can be resolved. As an example, we present here the dynamical anisotropy obtained by the electrical patterning of a film consisting of a self-assembled monolayer deposited on gold, covered with a phospholipid hemimembrane.

Nanoplasmonics-enhanced label-free imaging of endothelial cell monolayer integrity.

Surface plasmon resonance imaging (SPRI) is a powerful label-free imaging modality for the analysis of morphological dynamics in cell monolayers. However, classical plasmonic imaging systems have relatively poor spatial resolution along one axis due to the plasmon mode attenuation distance (tens of µm, typically), which significantly limits their ability to resolve subcellular structures. We address this limitation by adding an array of nanostructures onto the metal sensing surface (25 nm thick, 200 nm width, 400 nm period grating) to couple localized plasmons with propagating plasmons, thereby reducing attenuation length and commensurately increasing spatial imaging resolution, without significant loss of sensitivity or image contrast. In this work, experimental results obtained with both conventional unstructured and nanostructured gold film SPRI sensor chips show a clear gain in spatial resolution achieved with surface nanostructuring. The work demonstrates the ability of the nanostructured SPRI chips to resolve fine morphological detail (intercellular gaps) in experiments monitoring changes in endothelial cell monolayer integrity following the activation of the cell surface protease-activated receptor 1 (PAR1) by thrombin. In particular, the nanostructured chips reveal the persistence of small intercellular gaps (<5 µm2) well after apparent recovery of cell monolayer integrity as determined by conventional unstructured surface based SPRI. This new high spatial resolution plasmonic imaging technique uses low-cost and reusable patterned substrates and is likely to find applications in cell biology and pharmacology by allowing label-free quantification of minute cell morphological activities associated with receptor dependent intracellular signaling activity.

Spectroscopic ultrahigh-resolution full-field optical coherence microscopy.

We have developed a full-field optical coherence microscopy system providing intensity-based tomographic images and spectroscopic information with ultrahigh spatial resolution. Local measurements of the backscattered light spectrum center of mass are achieved through short-time Fourier analysis of a stack of en face interferometric images acquired with a Linnik-type microscope. Using a halogen lamp as an illumination source enables us to achieve spectroscopic imaging over a wavelength range from 600 to 900 nm with a spatial resolution of approximately 1 microm. Absorption measurements of a colored gel are reported as a validation of the technique. Enhancement of tissue imaging contrast is demonstrated by imaging a Xenopus Laevis (African frog) tadpole ex vivo.

Simultaneous dual-band ultra-high resolution full-field optical coherence tomography.

Ultrahigh-resolution full-field optical coherence tomography (FF-OCT) is demonstrated in the 800 nm and 1200 nm wavelength regions simultaneously using a Silicon-based (Si) CCD camera and an Indium Gallium Arsenide (InGaAs) camera as area detectors and a halogen lamp as illumination source. The FF-OCT setup is optimized to support the two broad spectral bands in parallel, achieving a detection sensitivity of approximately 90 dB and a micrometer-scale resolution in the three directions. Images of ex vivo biological tissues are presented (rabbit trachea and Xenopus laevis tadpole) with an increase in penetration depth at 1200 nm. A color image representation is applied to fuse both images and enhance spectroscopic property visualization.

Cybercare 2.0: meeting the challenge of the global burden of disease in 2030.

In this paper, we propose to advance and transform today's healthcare system using a model of networked health care called Cybercare. Cybercare means "health care in cyberspace" - for example, doctors consulting with patients via videoconferencing across a distributed network; or patients receiving care locally - in neighborhoods, "minute clinics," and homes - using information technologies such as telemedicine, smartphones, and wearable sensors to link to tertiary medical specialists. This model contrasts with traditional health care, in which patients travel (often a great distance) to receive care from providers in a central hospital. The Cybercare model shifts health care provision from hospital to home; from specialist to generalist; and from treatment to prevention. Cybercare employs advanced technology to deliver services efficiently across the distributed network - for example, using telemedicine, wearable sensors and cell phones to link patients to specialists and upload their medical data in near-real time; using information technology (IT) to rapidly detect, track, and contain the spread of a global pandemic; or using cell phones to manage medical care in a disaster situation. Cybercare uses seven "pillars" of technology to provide medical care: genomics; telemedicine; robotics; simulation, including virtual and augmented reality; artificial intelligence (AI), including intelligent agents; the electronic medical record (EMR); and smartphones. All these technologies are evolving and blending. The technologies are integrated functionally because they underlie the Cybercare network, and/or form part of the care for patients using that distributed network. Moving health care provision to a networked, distributed model will save money, improve outcomes, facilitate access, improve security, increase patient and provider satisfaction, and may mitigate the international global burden of disease. In this paper we discuss how Cybercare is being implemented now, and envision its growth by 2030.

Parallel Structured Mesh Generation with Disparity Maps by GPU Implementation.

The goal of structured mesh is to generate a compressed representation of the 3D surface, where near objects are provided with more details than objects far from the camera, according to the disparity map. The solution is based on the Kohonens Self-Organizing Map algorithm for the benefits of its ability to generate a topological map according to a probability distribution and its potential to be a natural massive parallel algorithm. The disparity map, which stands for a density distribution that reflects the proximity of objects to the camera, is partitioned into an appropriate number of cell units, in such a way that each cell is associated to a processing unit and responsible of a certain area of the plane. The advantage of the proposed model is that it is decentralized and based on data decomposition. The required processing units and memory are with linearly increasing relationship to the problem size. Experimental results show that our GPU implementation is able to provide near real-time performance with small size disparity maps and the running time increases in a linear way with a very weak increasing coefficient. The proposed method is suitable to deal with large scale problems in a massively parallel way.

Confocal thermal-lens microscope.

The use of a confocal detection scheme in a dual-beam thermal-lens microscope is presented. The scheme allows the measurement of absorption factors down to 1.2 x 10(-7) in a 0.35-microm3 volume by use of a heating laser power of 100 mW incident upon the sample. Results are presented that prove that a 450-nm axial resolution is possible when a 1.2 water immersion objective lens with a N.A. of 1.2 is used.

Full-field birefringence imaging by thermal-light polarization-sensitive optical coherence tomography. I. Theory.

A method for measuring birefringence by use of thermal-light polarization-sensitive optical coherence tomography is presented. The use of thermal light brings to polarization-sensitive optical coherence tomography a resolution in the micrometer range in three dimensions. The instrument is based on a Linnik interference microscope and makes use of achromatic quarter-wave plates. A mathematical representation of the instrument is presented here, and the detection scheme is described, together with a discussion of the validity domain of the equations used to evaluate the birefringence in the presence of white-light illumination.