Conception d'un laboratoire sur fibre pour l'analyse in vivo

International audienceThis work presents the proof of concept of the detection of optical index changes by surface plasmon resonance (SPR) thanks to optical fiber bundles. This proof of concept is the first essential step for the future design of a lab on fiber tool dedicated to molecular analysis for endoscopic diagnosis. Our approach is based on nanotextured optical fiber bundles comprising several thousands of individual optical fibers. These nanostructures were coated by a thin gold layer in order to exhibit interesting optical properties like SPR. The sensitivity of the bundle to optical index changes and the detection limit were measured in retro-reflection. We performed numerical simulations in order to enhance these performances based on an optimization of the fiber end geometry and the gold coating thickness. We finally obtained a detection limit of 10-4 refractive index unit, which is fully compatible with the detection of biological interactions involving large proteins or bacteria. </p

Peptide-protein microarrays and surface plasmon resonance detection: biosensors for versatile biomolecular interaction analysis.

International audienceBiosensors in microarray format provide promising tools for high-throughput analyses of complex samples. Although they are able to detect, quantify and characterize a multitude of compounds, most of the available devices are specialized in the analysis of one type of interaction, limiting their application to a define area. The aim of our work was to develop and characterize versatile protein (or peptide) microarrays suitable for the simultaneous analysis of a large panel of biological interactions. Our system involved a simple procedure to immobilized proteins or peptides, based on pyrrole electropolymerization, and ligand binding was detected by imaging the surface plasmon resonance. We demonstrated its suitability in three different contexts, i.e. humoral response characterization, ion binding analysis and cell detection. This work evidences the potentiality of this approach which allows multiparametric, high-throughput and label-free analysis of biological samples suitable for the detection of compounds as various as proteins, ions or cells and the characterization of their interaction with peptides or proteins

Chauffage par effet plasmon pour la désorption localisée de cellules capturées sur une biopuce

International audienceThis work proposes a miniaturized system able to perform multiple cell capture followed by cell-type selective release from a biochip surface. Unlabeled lymphocytes are first specifically captured onto a DNA array by antibody-DNA conjugates. The immobilized cells are subsequently released under spatio-temporal control within local heating generated by intense Surface Plasmon Resonance (SPR) produced by laser illumination

Relationship between humoral response against hepatitis C virus and disease overcome

International audienceConclusionHumoral response against hepatitis C virus linear epitopes is partly modified according to the disease state. This study highlights the importance of considering relative quantities of antibodies with different specificities rather than the amount of each antibody.Hepatitis C virus infection leads to liver disease whose severity can range from mild to serious lifelong illness. However the parameters involved in the evolution of the disease are still unknown. Among other factors, the virus-elicited antibody profile is suspected to play a role in the outcome of the disease. Analysis of the relationship between anti-virus antibodies and disease state requires the analysis of a large number of serums from patients (hepatitis C virus+) and of epitopes from the viral proteins. Such a study would benefit from microarray-based screening systems that are appropriate for high-throughput assays.We used a method combining peptide chips and surface plasmon resonance imaging previously shown to be suitable for analyzing complex mediums and detecting peptide-protein interactions. 56 peptides covering the entire viral proteome were grafted on chips and their interaction with antibodies present in the 68 injected serums from infected and non-infected donors was measured. Statistical analyses were conducted to determine a possible relationship between antibodies (specificity and amount) and disease states.A good discrimination between infected and non-infected donors validated our approach, and several correlations between antibodies profiles and clinical parameters have been identified. In particular, we demonstrated that ratios between particular antibodies levels allow for accurate discrimination of patients according to their pathologic states

Road Map for Nanocrystal Based Infrared Photodetectors

Infrared (IR) sensors based on epitaxially grown semiconductors face two main challenges which are their prohibitive cost and the difficulty to rise the operating temperature. The quest for alternative technologies which will tackle these two difficulties requires the development of new IR active materials. Over the past decade, significant progresses have been achieved. In this perspective, we summarize the current state of the art relative to nanocrystal based IR sensing and stress the main materials, devices and industrial challenges which will have to be addressed over the 5 next years

A DNA biochip for on-the-spot multiplexed pathogen identification

Miniaturized integrated DNA analysis systems have largely been based on a multi-chamber design with microfluidic control to process the sample sequentially from one module to another. This microchip design in connection with optics involved hinders the deployment of this technology for point-of-care applications. In this work, we demonstrate the implementation of sample preparation, DNA amplification, and electrochemical detection in a single silicon and glass-based microchamber and its application for the multiplexed detection of Escherichia coli and Bacillus subtilis cells. The microdevice has a thin-film heater and temperature sensor patterned on the silicon substrate. An array of indium tin oxide (ITO) electrodes was constructed within the microchamber as the transduction element. Oligonucleotide probes specific to the target amplicons are individually positioned at each ITO surface by electrochemical copolymerization of pyrrole and pyrrole−probe conjugate. These immobilized probes were stable to the thermal cycling process and were highly selective. The DNA-based identification of the two model pathogens involved a number of steps including a thermal lysis step, magnetic particle-based isolation of the target genomes, asymmetric PCR, and electrochemical sequence-specific detection using silver-enhanced gold nanoparticles. The microchamber platform described here offers a cost-effective and sample-to-answer technology for on-site monitoring of multiple pathogens

A DNA biochip for on-the-spot multiplexed pathogen identification

Miniaturized integrated DNA analysis systems have largely been based on a multi-chamber design with microfluidic control to process the sample sequentially from one module to another. This microchip design in connection with optics involved hinders the deployment of this technology for point-of-care applications. In this work, we demonstrate the implementation of sample preparation, DNA amplification, and electrochemical detection in a single silicon and glass-based microchamber and its application for the multiplexed detection of Escherichia coli and Bacillus subtilis cells. The microdevice has a thin-film heater and temperature sensor patterned on the silicon substrate. An array of indium tin oxide (ITO) electrodes was constructed within the microchamber as the transduction element. Oligonucleotide probes specific to the target amplicons are individually positioned at each ITO surface by electrochemical copolymerization of pyrrole and pyrrole−probe conjugate. These immobilized probes were stable to the thermal cycling process and were highly selective. The DNA-based identification of the two model pathogens involved a number of steps including a thermal lysis step, magnetic particle-based isolation of the target genomes, asymmetric PCR, and electrochemical sequence-specific detection using silver-enhanced gold nanoparticles. The microchamber platform described here offers a cost-effective and sample-to-answer technology for on-site monitoring of multiple pathogens