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We report an optofluidic hybrid silicon-polymer planar ring resonator with integrated microfluidic channels for efficient liquid delivery. The device features a planar architecture of intersecting liquid-core waveguides and microfluidic channels. A low-loss integration of microfluidic channels is accomplished by exploiting the interference pattern created by the self-imaging effect in the multimode interference-based coupler waveguides. Numerical simulations have been performed in order to minimize the propagation losses along the ring loop caused by the integration of microfluidic channels. The device has been fabricated and optically characterized by measuring the quality factor, obtaining a value of 4 × 103. This result is comparable with the quality factor of an optofluidic ring with the same optical layout but without integrated microfluidic channels, thus, confirming the suitability of the proposed approach for microfluidics integration in planar optofluidic design.
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BACKGROUND: Peptidoglycan is an essential component of the cell wall in all bacteria. In particular, the cell walls of Gram-positive bacteria are composed mostly of a thick layer of peptidoglycan. Its accessibility has important implications for their sensing in whole bacterial detection methodologies. Indeed, there is an urgent demand for rapid tests which can identify whole bacteria, e.g., directly at the point of care. OBJECTIVE: The aim of this work is to explore the suitability of RipA, a key cell division protein of M. tuberculosis, for whole cell biosensing of Gram-positive bacteria. METHODS: We here conducted Molecular Dynamics (MD) studies aimed at the understanding of the structural and dynamic features of active RipA and at the design of a suitable bioreceptor. Based on these studies, we engineered a RipA variant for covalent oriented immobilisation on golden surfaces and are able to bind peptidoglycan, albeit without degrading it. Surface Plasmon Resonance (SPR) was employed to check the ability of functionalized golden chips to recognize whole bacteria. RESULTS: MD analyses elucidated the structural details of the active form of RipA and suggested that this enzyme, once inactivated, presents a rigid and well-exposed peptidoglycan recognition cleft. We engineered RipA for proper oriented immobilisation on golden chips for SPR studies. Results show that once chemically coupled to a golden chip, the developed RipA-based bioreceptor is able to detect B. subtilis, used as a model in a concentration-dependent mode. CONCLUSION: Results highlight the potential of the engineered molecule to be integrated in the development of early warning biosensors for Gram-positive contamination in clinical diagnosis or food-borne infections.
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Proteínas Bacterianas , Endopeptidasas , Mycobacterium tuberculosis , Proteínas Bacterianas/metabolismo , Pared Celular/metabolismo , Endopeptidasas/metabolismo , Hidrolasas/metabolismo , Mycobacterium tuberculosis/metabolismo , Peptidoglicano/metabolismoRESUMEN
The present paper describes a compact point of care (POC) optical device for therapeutic drug monitoring (TDM). The core of the device is a disposable plastic chip where an immunoassay for the determination of immunosuppressants takes place. The chip is designed in order to have ten parallel microchannels allowing the simultaneous detection of more than one analyte with replicate measurements. The device is equipped with a microfluidic system, which provides sample mixing with the necessary chemicals and pumping samples, reagents and buffers into the measurement chip, and with integrated thin film amorphous silicon photodiodes for the fluorescence detection. Submicrometric fluorescent magnetic particles are used as support in the immunoassay in order to improve the efficiency of the assay. In particular, the magnetic feature is used to concentrate the antibody onto the sensing layer leading to a much faster implementation of the assay, while the fluorescent feature is used to increase the optical signal leading to a larger optical dynamic change and consequently a better sensitivity and a lower limit of detection. The design and development of the whole integrated optical device are here illustrated. In addition, detection of mycophenolic acid and cyclosporine A in spiked solutions and in microdialysate samples from patient blood with the implemented device are reported.
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Inmunosupresores , Dispositivos Ópticos , Humanos , Inmunoensayo , Microfluídica , SilicioRESUMEN
A full polymeric optofluidic Fabry-Perot (FP) resonator with a high quality factor (Q) is proposed and tested. The device is based on multilayer optical polymeric films that act as high reflectivity interference mirrors. The all-polymeric laminated structure avoids any deposition or etching process, simplifying the fabrication procedure while retaining a high quality optical surface. The measured quality factor of the FP resonator is 3.03×104, and the finesse is 91 around 700 nm. The refractometric sensing properties have been measured, and a sensitivity of 314 nm/RIU with a limit of detection of 2.55×10-5RIU could be achieved. The device exhibits a very high figure of merit of 1.36×104RIU-1 that is comparable with the performance attainable with sensors based on photonic crystal and whispering gallery mode resonators.
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In this paper, we report the study on a new protocol for the immobilization process of antigen/antibody assay on SU-8 layers by oxygen plasma treatment. Plasma treatments, at different plasma powers and for different duration times, are performed and their effects on immobilization efficiency are studied. The chemical properties and the surface morphology of SU-8 before and after the functionalization and immobilization of (IgG) are then verified by Raman spectroscopy and atomic force microscopy (AFM). An increase of the surface roughness of SU-8 layers is observed after the oxygen plasma treatment and an intensity variation of functional groups is also evidenced. To demonstrate the validity of the process the distribution of IgG immobilized on SU-8 surfaces is detected by fluorescence microscopy measurement after incubation with fluorescein isothiocyanate (FITC)-tagged anti-human IgG. An increase of the amount of the adsorbed protein of about 20% and a good repeatability on antigen/antibody distribution on the surface are detected for IgG on plasma treated substrates. Finally, label free measurements are performed by SU-8 optical ring resonators reaching detection limits of 0.86ngcm(-2). The proposed approach offers a smart protocol for IgG immobilization on SU-8 substrate that can be easily extended to different antigen/antibody assay and polymeric materials for the realization of high performance immunosensors.
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Anticuerpos/inmunología , Compuestos Epoxi/química , Inmunoensayo/instrumentación , Inmunoglobulina G/inmunología , Gases em Plasma/química , Polímeros/química , Resonancia por Plasmón de Superficie/instrumentación , Adsorción , Anticuerpos/análisis , Anticuerpos/química , Diseño de Equipo , Análisis de Falla de Equipo , Humanos , Inmunoglobulina G/análisis , Oxígeno/química , Unión Proteica , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
This publisher's note corrects the author listing of a publication.
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In this paper, we introduce a liquid core antiresonant reflecting optical waveguide (ARROW) as a novel optofluidic device that can be used to create innovative and highly functional microsensors. Liquid core ARROWs, with their dual ability to guide the light and the fluids in the same microchannel, have shown great potential as an optofluidic tool for quantitative spectroscopic analysis. ARROWs feature a planar architecture and, hence, are particularly attractive for chip scale integrated system. Step by step, several improvements have been made in recent years towards the implementation of these waveguides in a complete on-chip system for highly-sensitive detection down to the single molecule level. We review applications of liquid ARROWs for fluids sensing and discuss recent results and trends in the developments and applications of liquid ARROW in biomedical and biochemical research. The results outlined show that the strong light matter interaction occurring in the optofluidic channel of an ARROW and the versatility offered by the fabrication methods makes these waveguides a very promising building block for optofluidic sensor development.
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A micro flow cytometer with a single step 3D hydrodynamic flow focusing has been developed. The proposed design is capable to create a single-file particle stream that is self-aligned with an integrated optical fiber-based detection system, regardless of the flow rate ratio between the focusing and core liquids. The design approach provides the ability to adjust the stream size while keeping the position of the focused stream centered with respect to the focusing channel. The device has been fabricated by direct micro milling of PMMA sheets. Experimental validation of the hydrodynamic sheath focusing effect has been presented and sample stream with tuneable size from about 18 to 50 µm was measured. Flow cytometry measurements have been performed by using 10-23 µm fluorescent particles. From the analysis of the signals collected at each transit event we can confirm that the device was capable to align and measure microparticles with a good coefficient of variance.
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A hybrid silicon-poly(dimethysiloxane) (PDMS) optofluidic platform for lab-on-a-chip applications is proposed. A liquid-core waveguide with a self-aligned solid-core waveguide and a microfluidic device are integrated with a multilayer approach, resulting in a three-dimensional device assembly. The optofluidic layer was fabricated with a hybrid silicon-polymer technology, whereas the microfluidic layer was fabricated with a soft lithography technique. The combination of different materials and fabrication processes allows a modular approach, enabling both the benefits from the high optical quality achievable with silicon technology and the low cost of polymer processing. The proposed chip has been tested for fluorescence measurements on Cy5 water solutions, demonstrating the possibility to obtain a limit of detection of 2.5 nM.
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Optofluidics is a relatively young research field able to create a tight synergy between optics and micro/nano-fluidics. The high level of integration between fluidic and optical elements achievable by means of optofluidic approaches makes it possible to realize an innovative class of sensors, which have been demonstrated to have an improved sensitivity, adaptability and compactness. Many developments in this field have been made in the last years thanks to the availability of a new class of low cost materials and new technologies. This review describes the Italian state of art on optofluidic devices for sensing applications and offers a perspective for further future advances. We introduce the optofluidic concept and describe the advantages of merging photonic and fluidic elements, focusing on sensor developments for both environmental and biomedical monitoring.
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A novel spectroscopic sensor based on an optofluidic liquid jet waveguide is presented. In this device, a liquid jet waveguide is generated with the solution under analysis. This stream, exploiting total internal reflection, acts as an optical waveguide confining the autofluorescence light produced by chemical or biological samples when opportunely excited. Using a self-aligned configuration, the liquid jet is directly coupled with a multimode optical fiber collecting the fluorescence towards the detection system. Experimental measurements have been performed using an UV excitation source on water solutions containing representative water pollutants as aromatic hydrocarbons or bacteria showing very low limit of detection.
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Óptica y Fotónica/métodos , Espectrometría de Fluorescencia/métodos , Espectrofotometría Ultravioleta/métodos , Bacterias/aislamiento & purificación , Calibración , Límite de Detección , Naftalenos/análisis , Soluciones , Tolueno/análisis , Agua/químicaRESUMEN
An optofluidic water-jet waveguide on chip for fluorescence analysis is presented. A high-speed water stream produced by means of a microchannel acts at the same time as the solution to analyze and as the collecting optical waveguide. The absence of solid walls and the very optically smooth surface of the liquid stream permits a strong increase of the signal-to-noise ratio. The device layout allows a self-alignment direct coupling of a water-jet waveguide with a multimode optical fiber connected to the detector. The performances of the integrated system are tested on Cy5 water solutions. For a 4.5 mm long water-jet waveguide, the measured limit of detection (LOD) is 2.56 nM and filter-free detection is possible with an LOD of 6.11 nM.
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Rayos Láser , Microfluídica/instrumentación , Dispositivos Ópticos , Espectrometría de Fluorescencia/instrumentación , Resonancia por Plasmón de Superficie/instrumentación , Diseño de Equipo , Análisis de Falla de EquipoRESUMEN
We present an integrated tunable liquid-core/liquid-cladding (L2) optical fiber, based on a novel three-dimensional hydrodynamic focusing scheme that enables the production of a tunable circular liquid core located in the center of the channel, regardless of the flow-rate ratio of the cladding and core liquids.
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A high-visibility integrated optofluidic Mach-Zehnder interferometer based on liquid-core antiresonant reflecting optical waveguides is reported. The device's geometry has been optimized to minimize the intensity imbalance between the two arms for highly unbalanced Mach-Zehnder configurations. This results in a very compact device with a total length of only 2.5 mm and with required liquid volume of about 0.16 nl. High visibility is demonstrated for two interferometers corresponding to different sensing lengths. The devices have been optically characterized, and the measured interference fringes in the transmitted spectra show good agreement with the theoretical ones.
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In this work we describe the fabrication and the characterization of perfluorinated plastic-cladded optical fiber tapers. The heat-and-pull procedure has been used to fabricate symmetric tapers. Devices with different taper ratio have been produced and the repeatability of the process has been verified. The very low refractive indexes of the core-cladding perfluorinated polymers (n = 1.35-1.34) permit a strong enhancement of the evanescent wave power fraction in aqueous environments (n = 1.33), making them very attractive for evanescent wave sensing. The tapers have been characterized carrying out evanescent field absorbance measurements with different concentrations of methylene blue in water and fluorescence collection measurements in an aqueous solution containing Cy5 dye. A good sensitivity, tightly related to the low refractive index of the core-cladding materials and the geometrical profile, has been shown.