RESUMEN
A fast surface plasmon resonance (SPR) imaging biosensor system based on wavelength interrogation using an acousto-optic tunable filter (AOTF) and a white light laser is presented. The system combines the merits of a wide-dynamic detection range and high sensitivity offered by the spectral approach with multiplexed high-throughput data collection and a two-dimensional (2D) biosensor array. The key feature is the use of AOTF to realize wavelength scan from a white laser source and thus to achieve fast tracking of the SPR dip movement caused by target molecules binding to the sensor surface. Experimental results show that the system is capable of completing a SPR dip measurement within 0.35 s. To the best of our knowledge, this is the fastest time ever reported in the literature for imaging spectral interrogation. Based on a spectral window with a width of approximately 100 nm, a dynamic detection range and resolution of 4.63 × 10-2 refractive index unit (RIU) and 1.27 × 10-6 RIU achieved in a 2D-array sensor is reported here. The spectral SPR imaging sensor scheme has the capability of performing fast high-throughput detection of biomolecular interactions from 2D sensor arrays. The design has no mechanical moving parts, thus making the scheme completely solid-state.
RESUMEN
Imaging-based spectral surface plasmon resonance (λSPR) biosensing is predominantly limited by data throughput because of the multiplied data capacity emerging from 2-dimensional sensor array sites and the many data points required to produce an accurate measurement of the absorption dip. Here we present an adaptive feedback approach to address the data throughput issue in λSPR biosensing. A feedback loop constantly tracks the dip location while target-molecule binding occurs at the sensor surface. An adaptive window is then imposed to reduce the number of data points that each pixel has to capture without compromising measurement accuracy. Rapid wavelength scanning is performed with a liquid crystal tunable filter (LCTF). With the use of a feedback loop, our demonstration system can produce a dip measurement within 700ms, thus confirming that the reported λSPR approach is most suitable for real-time micro-array label-free biosensing applications.
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Técnicas Biosensibles , Resonancia por Plasmón de Superficie/métodosRESUMEN
Incorporating the temporal carrier technique with common-path spectral interferometry, we have successfully demonstrated an advanced surface plasmon resonance (SPR) biosensing system which achieves refractive index resolution (RIR) up to 2 × 10(-8) refractive index unit (RIU) over a wide dynamic range of 3 × 10(-2) RIU. While this is accomplished by optimizing the SPR differential phase sensing conditions with just a layer of gold, we managed to address the spectral phase discontinuity with a novel spectral-temporal phase measurement scheme. As the new optical setup supersedes its Michelson counterpart in term of simplicity, we believe that it is a significant contribution for practical SPR sensing applications.
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A wavelength-multiplexing phase-sensitive surface plasmon resonance (SPR) imaging sensor offering wide dynamic detection range and microarray capability is reported. Phase detection is accomplished by performing self-interference between the s- and p- polarizations within the signal beam. A liquid crystal tunable filter is used to sequentially select the SPR excitation wavelength from a white light source. This wavelength-multiplexing approach enables fast detection of the sensor's SPR phase response over a wide range of wavelengths, thereby covering literally any regions of interest within the SPR dip and thus maintaining the highest sensitivity point at all times. The phase-sensitive approach is particularly important for imaging SPR sensing applications because of its less stringent requirements for intensity signal-to-noise ratio, which also means the possibility of using uncooled modest resolution analog-to-digital conversion imaging devices. Experimental results demonstrate a resolution of 2.7×10(-7) RIU with a dynamic range of 0.0138 RIU.
Asunto(s)
Imagen Óptica/métodos , Resonancia por Plasmón de Superficie/métodos , Animales , Glicerol/química , Humanos , Inmunoglobulina G/inmunología , Agua/químicaRESUMEN
A novel differential phase detecting surface plasmon resonance (SPR) sensor based on white-light spectral interferometry is presented. Our proposed scheme employs a white-light source for SPR excitation and measures the corresponding SPR phase change at the optimized coupling wavelength with fixed angle of incidence across the visible spectrum. Compared to existing laser based phase detecting schemes, this system offers optimal sensitivity and extended dynamic range of measurement without any compromise in phase detection resolution. Results obtained from sodium chloride solutions indicate that the detection limit is 2.6×10â»7 RIU over a refractive index range of 10⻲ RIU, which is considerably wider than that achievable by existing laser based approach, thus making our scheme very attractive for practical SPR sensing applications.
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Interferometría/instrumentación , Análisis Espectral/instrumentación , Resonancia por Plasmón de Superficie/instrumentación , Transductores , Diseño de Equipo , Análisis de Falla de Equipo , LuzRESUMEN
Two-dimensional surface plasmon resonance (2D-SPR) imaging, which provides a real-time, sensitive, and high-throughput analysis of surface events in a two dimensional manner, is a valuable tool for studying biomolecular interactions and biochemical reactions without using any tag labels. The sensing principle of 2D-SPR includes angular, wavelength, and phase interrogation. In this chapter, the 2D-SPR imaging technique is applied for sensing a target microRNA by its corresponding oligonucleotide probes, with sequence complementarity, immobilized on the gold SPR sensing surface. However, the low SPR signal due to intrinsic properties such as low molecular weight and quantity (pico-nanomolar) of the microRNA in clinical samples limits the direct detection of microRNA. Therefore, we developed a biosensing technique known as MARS (MicroRNA-RNase-SPR) assay, which utilizes RNase H to digest the microRNA probes enzymatically for fast signal amplification, i.e., in order to increase both the SPR signal and readout speed without the need for pre-amplification of target cDNA by polymerase chain reaction (PCR). Practically, we targeted microRNA hsa-miR-29a-3p, whose signature correlates to influenza infection, for rapid screening of influenza A (H1N1) patients from throat swab samples.
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Técnicas Biosensibles/métodos , MicroARNs , Resonancia por Plasmón de Superficie/métodos , Especificidad de Anticuerpos , Técnicas Biosensibles/instrumentación , Humanos , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/diagnóstico , Gripe Humana/virología , MicroARNs/análisis , MicroARNs/genética , Resonancia por Plasmón de Superficie/instrumentación , Flujo de TrabajoRESUMEN
A lab-on-a-disc (LOAD) is a centrifugal microfluidic set-up based on centrifugal force without using micro-pumps to drive reagents and cells to various chambers through channels and valves for reactions. A LOAD coated with conductive transparent indium tin oxide (ITO) for thermal control was developed to screen allergy-blocking agents. When the acridine orange (AO)-loaded KU-812 human basophilic cells were activated in the LOAD by stimuli, AO trapped in the cytoplasmic granules was released externally as an allergic mediator mimetic to report degranulation. This response was monitored by fluorescence when the released AO in supernatant had been transferred, with a higher spinning speed, from the reaction chamber to detection chamber in the LOAD where AO reacted with exogenous DNA. We report here the principles of the system and an improved LOAD set-up with the ITO-coated glass resistive microheater to run assays at 37 °C. By using this platform, we demonstrate here for the first time that triptolide, an active ingredient from the Chinese medicine herb Tripterygium wilfordii Hook f., was able to suppress the fMLP-mediated degranulation in basophils. This serves as an example how LOADs can be used to screen agents to alleviate symptoms of allergy.
RESUMEN
A fast surface plasmon resonance (SPR) imaging biosensor system based on wavelength interrogation using a liquid crystal tunable filter (LCTF) is presented. The system combines the merits of wide-dynamic detection range offered by the spectral approach and multiplexed high-throughput data collection with a two-dimensional (2-D) biosensor array. The key feature of the reported scheme is a feedback loop that drives the LCTF to achieve fast tracking of the SPR dip movement caused by the binding of target molecules to the sensor surface. Experimental results show that the system is capable of completing an SPR dip measurement within 4 s. Based on using a spectral window of about 100 nm, the experimental dynamic detection range and refractive index resolution are 4.63×10?2??RIU and 5.87×10?6??RIU, respectively. As also demonstrated herein using 2-D microsensor arrays, among the spectral SPR sensors, the reported system is most suitable for multiplexed label-free biosensing applications.
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Técnicas Biosensibles/métodos , Procesamiento de Imagen Asistido por Computador/métodos , Resonancia por Plasmón de Superficie/métodos , Animales , Técnicas Biosensibles/instrumentación , Diseño de Equipo , Inmunoglobulina G/análisis , Cristales Líquidos , Análisis por Micromatrices , Relación Señal-Ruido , Resonancia por Plasmón de Superficie/instrumentaciónRESUMEN
Optical tweezers are a well-established tool for manipulating small objects. However, their integration with microfluidic devices often requires an objective lens. More importantly, trapping of non-transparent or optically sensitive targets is particularly challenging for optical tweezers. Here, for the first time, we present a photon-free trapping technique based on electro-thermally induced forces. We demonstrate that thermal-gradient-induced thermophoresis and thermal convection can lead to trapping of polystyrene spheres and live cells. While the subject of thermophoresis, particularly in the micro- and nano-scale, still remains to be fully explored, our experimental results have provided a reasonable explanation for the trapping effect. The so-called thermal tweezers, which can be readily fabricated by femtosecond laser writing, operate with low input power density and are highly versatile in terms of device configuration, thus rendering high potential for integration with microfluidic devices as well as lab-on-a-chip systems.
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Calor , Dispositivos Laboratorio en un Chip , Pinzas ÓpticasRESUMEN
We experimentally demonstrated the use of random plasmonic nano-islands for optical trapping and assembling of particles and live cells into highly organized pattern with low power density. The observed trapping effect is attributed to the net contribution due to near-field optical trapping force and long-range thermophoretic force, which overcomes the axial convective drag force, while the lateral convection pushes the target objects into the trapping zone. Our work provides a simple platform for on-chip optical manipulation of nano- and micro-sized objects, and may find applications in physical and life sciences.
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Allergies occur when a person's immune system mounts an abnormal response with or without IgE to a normally harmless substance called an allergen. The standard skin-prick test introduces suspected allergens into the skin with lancets in order to trigger allergic reactions. This test is annoying and sometimes life threatening. New tools such as lab-on-a-chip and lab-on-a-disc, which rely on microfabrication, are designed for allergy testing. These systems provide benefits such as short analysis times, enhanced sensitivity, simplified procedures, minimal consumption of sample and reagents and low cost. This article gives a summary of these systems. In particular, a cell-based assay detecting both the IgE- and non-IgE-type triggers through the study of degranulation in a centrifugal microfluidic system is highlighted.
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Alérgenos/análisis , Hipersensibilidad/diagnóstico , Inmunoglobulina E/análisis , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/instrumentación , Alérgenos/inmunología , Animales , Prueba de Desgranulación de los Basófilos/economía , Prueba de Desgranulación de los Basófilos/instrumentación , Prueba de Desgranulación de los Basófilos/métodos , Basófilos/inmunología , Basófilos/fisiología , Degranulación de la Célula , Diseño de Equipo , Humanos , Hipersensibilidad/inmunología , Inmunoglobulina E/inmunología , Dispositivos Laboratorio en un Chip/economía , Mastocitos/inmunología , Mastocitos/fisiología , Técnicas Analíticas Microfluídicas/economía , Técnicas Analíticas Microfluídicas/métodosRESUMEN
This report describes a novel diagnostic assay for rapid detection of the Panton-Valentine Leukocidin (PVL) toxin of methicillin-resistant Staphylococcus aureus (MRSA) utilizing resistive pulse sensing (RPS), loop-mediated isothermal DNA amplification (LAMP) in combination with gold nanoparticles (AuNPs). The PVL DNA from MRSA was specifically amplified by LAMP using four primers at one temperature (65 °C). The DNA products with biotin were then conjugated to a first AuNP1 (55±2 nm) through biotin-avidin binding. A second AuNP2 (30±1.5 nm) coated with a specific DNA probe hybridized with the LAMP DNA products at the loop region to enhance assay sensitivity and specificity, to generate supra-AuNP1-DNA-AuNP2 assemblies. Scanning electron microscopy confirmed the presence of these supra-assemblies. Using RPS, detection and quantitation of the agglomerated AuNPs were performed by a tunable fluidic nanopore sensor. The results demonstrate that the LAMP-based RPS sensor is sensitive and rapid for detecting the PVL DNA. This technique could achieve a limit of detection (LOD) up to about 500 copies of genomic DNA from the bacteria MRSA MW2 and the detection can be completed within two hours with a straightforward signal-to-readout setup. It is anticipated that this LAMP-based AuNP RPS may become an effective tool for MRSA detection and a potential platform in clinical laboratory to report the presence or absence of other types of infectious agents.
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Toxinas Bacterianas/análisis , Toxinas Bacterianas/genética , ADN Bacteriano/análisis , Exotoxinas/análisis , Exotoxinas/genética , Leucocidinas/análisis , Leucocidinas/genética , Staphylococcus aureus Resistente a Meticilina/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Secuencia de Bases , Sondas de ADN , Oro , Límite de Detección , Datos de Secuencia Molecular , Nanopartículas , Técnicas de Amplificación de Ácido Nucleico/instrumentación , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y EspecificidadRESUMEN
In this paper, we present a lab-in-a-droplet bioassay strategy for a centrifugal microfluidics or lab-on-a-disc (LOAD) platform with three important advancements including density difference pumping, power to disc and bidirectional flow control. First, with the water based bioassay droplets trapped in a micro-channel filled with mineral oil, centrifugal force due to the density difference between the water and oil phases actuates droplet movement while the oil based medium remains stationary. Second, electricity is coupled to the rotating disc through a split-core transformer, thus enabling on-chip real-time heating in selected areas as desired and wireless programmable functionality. Third, an inertial mechanical structure is proposed to achieve bidirectional flow control within the spinning disc. The droplets can move back and forth between two heaters upon changing the rotational speed. Our platform is an essential and versatile solution for bioassays such as those involving DNA amplification, where localized temperature cycling is required. Finally, without the loss of generality, we demonstrate the functionality of our platform by performing real-time polymerase chain reaction (RT-PCR) in a linear microchannel made with PTFE (Teflon) micro-tubing.
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Técnicas Analíticas Microfluídicas/instrumentación , Aceite Mineral/química , Centrifugación , ADN/análisis , Electricidad , Dispositivos Laboratorio en un Chip , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Temperatura , Agua/química , Tecnología InalámbricaRESUMEN
We introduce a novel wide dynamic range phase-sensitive surface plasmon resonance (SPR) biosensor based on differential spectral interferometry. Superseding conventional spectroscopic approach where only the SPR dip is monitored, our system acquires the spectral phase information of the entire electromagnetic field that undergoes SPR transformation. Since the SPR-induced phase change is highly wavelength specific with fixed incident angle, ultra-high sensitivity achievable through phase-sensitive detection, as reported herein, is maintained continuously across the spectral domain in response to refractive index changes. Our system has demonstrated a detection limit of 2.2×10(-7) in terms of refractive index unit (RIU) using standard single-layer gold surface. In terms of biosensing performance, the estimated detection sensitivity obtained from bovine serum albumin (BSA) antibody-antigen binding experiments is 0.5 ng ml(-1).
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Técnicas Biosensibles/métodos , Espectroscopía Dieléctrica/métodos , Interferometría/métodos , Resonancia por Plasmón de Superficie/métodos , Animales , Reacciones Antígeno-Anticuerpo , Técnicas Biosensibles/instrumentación , Técnicas Biosensibles/estadística & datos numéricos , Bovinos , Espectroscopía Dieléctrica/instrumentación , Oro , Técnicas In Vitro , Interferometría/instrumentación , Límite de Detección , Refractometría , Albúmina Sérica Bovina/inmunología , Albúmina Sérica Bovina/metabolismo , Resonancia por Plasmón de Superficie/instrumentación , Resonancia por Plasmón de Superficie/estadística & datos numéricosRESUMEN
We present a new technique based on the spectral characteristics associated with the surface plasmon resonance (SPR) effect for studying lubricants in elastohydrodynamic (EHD) dimples. The pressure inside the EHD dimple causes a localized change of the refractive index (RI) of the entrapped lubricant. This also results in a shift in the spectral SPR absorption dip. By monitoring the color changes within the SPR image, one can obtain a direct measurement of the RI of the entrapped lubricant, from which the pressure distribution within the elastohydrodynamic lubrication (EHL) dimple can be deduced by means of a predetermined relation of pressure and RI of the tested lubricant. Dimples formed with the lubricants PB 2400 and H 1900 were studied in our experiments. Because SPR is sensitive only to the RI variation within a thin region (approximately one wavelength) close to the sensor's surface, the new technique does not require any measurement of the absolute film thickness of the lubricant. This is much simpler than the existing two-beam interferometric technique for measuring the RI of lubricants in EHD dimples, which requires simultaneous measurements of optical film thickness by use of two beams of different angles of incidence. In light of this advantage we anticipate that the new technique can be applied to pressure field mapping in highly loaded rolling and sliding EHL contacts.