Sensitivity enhancement of spectral surface plasmon resonance biosensors for the analysis of protein arrays

A novel method for sensitivity enhancement of spectral surface plasmon resonance (SPR) biosensors was presented by reducing the refractive index of the sensing prism in the analysis of protein arrays. Sensitivity of spectral SPR biosensors with two different prisms (BK-7, fused silica) was analyzed by net shifts of resonance wavelength for specific interactions of GST–GTPase binding domain of p21-activated kinase-1 and anti-GST on a mixed thiol surface. Sensitivity was modulated by the refractive index of the sensing prism of the spectral SPR biosensors with the same incidence angle. The sensitivity of a spectral SPR biosensor with a fused silica prism was 1.6 times higher than that with a BK-7 prism at the same incidence angle of 46.2°. This result was interpreted by increment of the penetration depth correlated with evanescent field intensity at the metal/dielectric interface. Therefore, it is suggested that sensitivity enhancement is readily achieved by reducing the refractive index of the sensing prism of spectral SPR biosensors to be operated at long wavelength ranges for the analysis of protein arrays.     

Analysis of protein interactions on protein arrays by a wavelength interrogation-based surface plasmon resonance biosensor

We have investigated whether surface plasmon resonance (SPR) sensors based on the wavelength interrogation are able to analyze protein interactions on protein arrays. The spectral SPR sensor was self-constructed and its detection limit, expressed as the minimal refractive index variation, was calculated to be 6.6x10(-5) with the signal fluctuation of 1.0x10(-5). The protein array surface was modified by a mixed thiol monolayer to immobilize proteins. Protein arrays were analyzed by the line-scanning mode of the SPR sensor, which scanned every 100 microm along the central line of array spots and the scanned results were presented by color spectra from blue to red. Glutathione S-transferase (GST)-rac1 caused a concentration-dependent increase of SPR wavelength shift on protein arrays. The surface structure of the protein arrays was analyzed by atomic force microscopy. Specific interactions of antigens with antibodies were analyzed on the protein arrays by using three antibodies and eight proteins. These results suggest that the wavelength interrogation-based SPR sensor can be used as the biosensor for the high-throughput analysis of protein interactions on protein arrays.     

Application of chromogenic reagents in surface plasmon resonance (SPR)

In this paper, a new simple approach for sensitivity optimization in surface plasmon resonance (SPR) chemosensors based on colorimetric ligands is presented. A new design of SPR sensor with tunable analytical wavelength (lambda(SPR)) was constructed for this purpose, to perform studies on the ligand absorbance spectra related sensitivity enhancement. Unlike commercial SPR sensors which operate at one lambda(SPR), the new device can be used for sensitivity analysis at selected lambda(SPR) in the range 550-750 nm, offering the possibility to identify the highest sensitivity lambda(SPR) in regard to the spectral changes of the selected ligand. Measurements can be easily done in ligand bulk solutions without immobilization steps. Sensitivity enhancement analysis and optimization of lambda(SPR) on chromogenic reagents with hypsochromic shift in their absorption spectra are demonstrated in this contribution. Optimal selection of analytical wavelength, set at the absorbance peak of chromogenic reagent Eriochrome Black T (EBT) was observed to result in up to two times increased SPR sensitivity to Cd(2+) compared to wavelengths selected in other parts of the ligand absorbance spectra, with a limit of detection (LOD) 0.2 ppm. The sensitivity enhancement at optimal lambda(SPR) was observed to be related to increased refractive index (n), drop in extinction coefficient (alpha) and simultaneous hypsochromic shift of the EBT absorbance spectra causing the lambda(SPR) to match the absorbance peak shoulder.     

A miniaturized germanium-doped silicon dioxide-based surface plasmon resonance waveguide sensor for immunoassay detection

A surface plasmon resonance (SPR) waveguide immunosensor fabricated by germanium-doped silicon dioxide was investigated in this study. The designed waveguide sensor consisted of a 10 microm SiO(2) substrate layer (n=1.469), a 10 microm Ge-SiO(2) channel guide (n=1.492) and a 50 nm gold film layer for immobilization of biomolecules and SPR signal detection. The resultant spectral signal was measured by a portable spectrophotometer, where the sensor was aligned by a custom-designed micro-positioner. The results of the glycerol calibration standards showed that the resonance wavelength shifted from 628 to 758 nm due to changes of refractive index from 1.36 to 1.418. Flow-through immunoassay on waveguide sensors also showed the interactions of protein A, monoclonal antibody (mAb ALV-J) and avian leucosis virus (ALVs) resulted in wavelength shifting of 4.17, 3.03 and 2.18 nm, respectively. The SPR dynamic interaction could also be demonstrated successfully in 4 min as the sensor was integrated with a lateral flow nitrocellulose strip. These results suggest that SPR detection could be carried out on designed waveguide sensor, and the integration of nitrocellulose strip for sample filtering and fluid carrier would facilitate applications in point-of-care portable system.     

Optimization of Surface Plasmon Resonance Biosensor with Ag/Au Multilayer Structure and Fiber-Optic Miniaturization

In this paper, we report a novel wavelength interrogation-based surface plasmon resonance (SPR) system, in which a film of three Ag layers and three Au layers are alternately deposited on a Kretschmann configuration as sensing element. This multilayer film shows higher sensitivity for refractive index (RI) measurement by comparing with single Au layer structure, which is consistent with its theoretical calculation. A sensitivity range of 2056–5893 nm/RIU can be achieved, which is comparable to RI sensitivities of other wavelength-modulated SPR sensors. Compared with Ag film, this Ag/Au multilayer arrangement offers anti-oxidant protection. This SPR biosensor based on a cost-effective Ag/Au multilayer structure is applicable to the real-time detection of specific interactions and dissociation of low protein concentrations. To extend the application of this highly-sensitive metal film device, we integrated this concept on an optical fiber. The range of RI sensitivities with Ag/Au multilayer was 1847–3309 nm/RIU. This miniaturized Ag/Au multilayer-based fiber optic sensor has a broad application in chemical and biological sensing.     

Comparison of Gold and Silver/Gold Bimetallic Surface for Highly Sensitive Near-infrared SPR Sensor at 1550 nm

A large majority of surface plasmon resonance (SPR) sensors reported in the literature are designed to operate in the visible electromagnetic spectrum. However, the near-infrared, particularly at the telecommunications wavelength of 1550 nm, is also especially attractive for SPR sensing applications. In fact, SPR sensors operating in this region benefit from narrower resonance and deeper field penetration. In this paper, we report a theoretical and experimental study of an SPR sensor operating at a fixed wavelength of 1550 nm. The influence of the choice of metals and the interrogation methods on the sensitivity of the resulting SPR sensor is investigated. Two types of sensor chips (simple gold (Au) and bimetallic silver/Au structure) and three interrogation methods (monitoring of the position of the reflectivity minimum, the position of the centroid, and the intensity evolution of the reflectivity) are examined. We show that a refractive index resolution of 2.7?×?10^?6 refractive index unit can be easily obtained, and with further optimization of the measurement system, the ultimate limit of detection is expected to be even lowered. Therefore, the approach discussed here already shows a promising potential for highly sensitive SPR sensors.     

Analysis of protein interactions on protein arrays by a novel spectral surface plasmon resonance imaging

We presented a novel surface plasmon resonance (SPR) imaging method for analysis of protein arrays based on a wavelength interrogation-based SPR biosensor. The spectral imaging was performed by the combination of position control and resonance wavelengths calculated from SPR reflectivity spectra. The imaging method was evaluated by analyzing interactions of glutathione S-transferase-fusion proteins with their antibodies. Antigen-antibody interactions were successfully analyzed on glutathione S-transferase-fusion protein arrays by using the spectral imaging method, and the results were confirmed by a parallel analysis using a previously used spectral SPR biosensor based on wavelength interrogation. Specific binding of anti-Rac1 and anti-RhoA to Rac1 and RhoA on the protein arrays was qualitatively and quantitatively analyzed by the spectral SPR imaging. Thus, it was suggested that the novel spectral SPR imaging was a useful tool for the high-throughput analysis of protein-protein interactions on protein arrays.     

Metallic Nanowire Array–Polymer Hybrid Film for Surface Plasmon Resonance Sensitivity Enhancement and Spectral Range Enlargement

In this paper, the coupling interaction is investigated between a metallic nanowire array and a metal film under the Kretschmann condition. The plasmonic multilayer is composed of a metallic nanowire array embedded in a polymer layer positioned above a metal film, exploiting the classical surface plasmon resonance (SPR) configuration. We analyze the influence of various structural parameters of the metallic nanowire array on the SPR spectrum of thin metal film. The results show that the coupling interactions of nanowires with the metal film can greatly affect SPR resonance wavelength and increase SPR sensitivity. The coupling strength of metallic nanowire array and metal film also impacts resonance wavelength, which can be used to adjust SPR range but have little effect on its sensitivity. The results are confirmed using a dipole coupling resonance model of metallic nanowire. We demonstrated that this nanostructured hybrid structure can be used for high sensitivity SPR monitoring in a large spectral range, which is important for advanced SPR measurement including fiber-optic SPR sensing technology.     

Highly Sensitive SPR Sensor Based on D-shaped Photonic Crystal Fiber Coated with Indium Tin Oxide at Near-Infrared Wavelength

A surface plasmon resonance (SPR) sensor based on D-shaped photonic crystal fiber (PCF) coated with indium tin oxide (ITO) film is proposed and numerically investigated. Thanks to the adjustable complex refractive index of ITO, the sensor can be operated in the near-infrared (NIR) region. The wavelength sensitivity, amplitude sensitivity, and phase sensitivity are investigated with different fiber structure parameters. Simulation results show that ～6000 nm/refractive index unit (RIU), ～148/RIU, and ～1.2?×?10⁶ deg/RIU/cm sensitivity can be achieved for wavelength interrogation, amplitude interrogation, and phase interrogation, respectively, when the environment refractive index varies between 1.30 and 1.31. It is noted that the wavelength sensitivity and phase sensitivity are more pronounced with larger refractive index. The proposed SPR sensor can be used in various applications, including medicine, environment, and large-scale targets detection.     

Determination of oxygen saturation and hematocrit of flowing human blood using two different spectrally resolving sensors.

The blood parameters oxygen saturation and hematocrit were determined by two different spectral sensors using reflectance spectra from 550 to 900 nm and partial transmission spectra centered at 660 nm. The spectra were analyzed by the method of partial least squares. One sensor consists of a miniature integrating sphere, while the other was fiber-guided. The results show that the geometry of the sensors and different blood flows do not influence the spectral analysis significantly. Independent of the sensor geometry, both hematocrit and oxygen saturation could be determined with an absolute predicted root mean square error of less than 3%. Furthermore, the analysis showed that hematocrit prediction requires eight wavelength regions and oxygen saturation prediction requires four wavelength regions using reflectance spectroscopy. This implies that if the measurement is restricted to reflectance, a spectrometer is indispensable for determining both blood parameters. Hematocrit determination could be improved using reflectance measurements in combination with transmission.     

Tunable Surface Plasmon Resonance Sensor Based on Photonic Crystal Fiber Filled with Gold Nanoshells

We present a photonic crystal fiber (PCF)-based surface plasmon resonance (SPR) sensor, whose operating wavelength range is tunable. Gold nanoshells, consisting of silica cores coated with thin gold shells, are designed to be the functional material of the sensor because of their attractive optical properties. It is demonstrated that the resonant wavelength of the sensor can be precisely tuned in a broad range, 660 nm to 3.1 μm, across the visible and near-infrared regions of the spectrum by varying the diameter of the core and the thickness of the shell. Furthermore, the effects of structural parameters of the sensor on the sensing properties are systematically analyzed and discussed based on the numerical simulations. It is observed that a high spectral sensitivity of 4111.4 nm/RIU with the resolution of 2.45 × 10^?5 RIU can be achieved in the sensing range of 1.33–1.38. These features make the sensor of great importance for a wide range of applications, especially in biosensing.     

Spectral Phase Shift of Surface Plasmon Resonance in the Kretschmann Configuration: Theory and Experiment

The spectral phase shift of surface plasmon resonance (SPR) in the Kretschmann configuration is modeled for aqueous solutions of NaCl (analytes) and an SPR structure consisting of gold and chromium layers deposited on an SF10 glass slide. Using the material dispersion of the SPR structure and the analyte, the SPR phase shift, its spectral derivative, and the spectral dependence of the ratio of the reflectances of p- and s-polarized waves are determined for aqueous solutions of NaCl when the concentration of NaCl in water and the refractive index range from 0 to 10 weight percent (wt%) and from 1.3334 to 1.3515 RIU, respectively. In addition, theoretical modeling is accompanied by experiment and the position of a sharp maximum in the measured spectral derivative of the SPR phase shift changes in a range from 596 to 626 nm. From the measurements, a sensitivity to concentration of 3.83 nm/wt% and a detection limit of 7.3 × 10^?7 RIU at a wavelength of 612.36 nm are obtained, and very good agreement between theory and experiment is confirmed.     

High-Sensitivity Refractive Index Sensor Based on D-Shaped Photonic Crystal Fiber with Rectangular Lattice and Nanoscale Gold Film

We propose and investigate a D-shaped photonic fiber refractive index sensor with rectangular lattice based on surface plasmon resonance. In such sensor, the nanoscale gold metal film is deposited on the flat surface where it is side polished. Numerical results show that the average sensitivity of Au-metalized surface plasmon resonance (SPR) sensor could reach as high as 8,129 nm/refractive index unit (RIU) in the dynamic index range from 1.35 to 1.41 as well as 2,000 nm/RIU from 1.33 to 1.35. Compared to conventional Au-metalized SPR sensors, the performance of our device is obviously better, and the production process is greatly simplified.     

Temperature Sensor Based on Hollow Fiber Filled with Graphene-Ag Composite Nanowire and Liquid

A temperature sensor based on hollow fiber (HF) filled with graphene-Ag composite nanowire and liquid is presented. The coupling properties and sensing performance are numerically analyzed by finite element method using wavelength and amplitude interrogations. Results show that the sensor exhibiting strong birefringence with x-polarized peak provides much higher sensitivities and better signal-to-noise ratio (SNR) than y-polarized, which is more suitable for temperature detection. The graphene-Ag composite nanowire can not only solve the oxidation problem but also avoid the metal coating. Moreover, it provides better performance than other similar works like Au-Ag nanowire-filled, Au nanowire-filled, and Ag nanowire-filled sensors. Contrary to the blue shift of traditional SPR temperature sensors, the resonance peak shifts to the longer wavelength in our device when temperature increases and the high sensitivity 9.44 nm/ °C is obtained. The influences of nanowire diameter, grapheme-layer thickness on the designed sensor, are also investigated. This work can provide a reference for developing a high sensitivity, real-time, remote sensing, and distributed temperature SPR sensor.     

An optical biosensor for monitoring recombinant proteins in process media.

This paper describes the construction of a sensor for the direct monitoring of a recombinant protein, the human insulin analogue (MI3). The surface plasmon resonance (SPR) sensor incorporates an immobilised, sterilisable affinity-ligand that has been designed to bind to MI3. In practice, gold SPR devices were fabricated with; a 2D assembly of ethanethiol-modified ligand, a 2D mixed-assembly of ethanethiol-modified ligand and mercaptoethanol, a 3D coating of ligand-modified terminal-thiolated poly(vinyl)alcohol (PVA) or a 3D hydrogel of dextran coupled to a self-assembled monolayer (SAM) of mercaptohexaneundecanl-ol. Routine measurement of the concentration MI3 in the concentration range 1-100 mg/l in pilot-scale samples of crude fermentation broth have been achieved with high sensitivity levels and a high signal-to-noise ratio. Analysis can be achieved within < 10 min with the active surface being regenerable for at least 60 cycles over a 6 month period. The coupling of a robust, sterilisable and highly-selective sensor-coating with suitable transducer technologies promises to deliver sensors that are capable of direct in situ monitoring of biopharmaceuticals in industrial bioprocesses.     

Reaction of cyanide with cytochrome aa3 in isolated perfused rat head in situ.

The reaction of cyanide with cytochrome aa3 in intact mitochondria is known to differ significantly from the reaction with the isolated enzyme. To examine the cyanide reaction with cytochrome aa3 in situ, we studied the spectral characteristics and the reaction kinetics of cyanide with reduced brain cytochrome aa3 in an isolated perfused rat head preparation. Anaesthetized rats underwent bilateral carotid-arterial cannulation. The head (skull intact, muscle removed) was perfused with a crystalloid solution containing Na2S2O4, and the animal was then decapitated. By means of reflectance spectrophotometry the reaction of cyanide with cytochrome aa3 was continuously monitored with the use of the 590 nm-575 nm, 610 nm-575 nm and 590 nm-610 nm wavelength pairs. We found that: the kinetics of the absorbance change at 590 nm and 610 nm were similar, with almost identical apparent rate constants, suggesting that these spectral changes are the results of the formation of a single complex; the difference spectrum obtained on addition of cyanide to the fully reduced preparation showed a peak at 588 nm and a trough at 610 nm, consistent with spectral characteristics of the cyanide-ferrocytochrome aa3 complex in isolated enzyme and isolated mitochondria in vitro; this observation underscores the accuracy of monitoring the effects of inhibitors of mitochondrial function on cytochrome redox reactions in situ; the half-maximal (K0.5) effect was approx. 50 microM, significantly lower than that in vitro. The lower apparent K0.5 for cyanide in this preparation in situ may be due to a difference in the pH of the two systems. This approach provides the means to study the inhibitors of mitochondrial function in intact brain under a physiological environment.     

Effects of light wavelength on growth and survival rate in juvenile Pacific bluefin tuna, Thunnus orientalis

The spectral sensitivity of the fish and the suitable light wavelength range for survival and growth performance of juvenile Pacific bluefin tuna (PBT) were investigated. The spectral sensitivity peak of PBT under photopic condition was observed between 449 and 503 nm, which corresponded to their natural habitat. The fish were reared in tanks irradiated continuously with 4 kinds of light emitting diodes (LEDs). The maximum wavelength of LEDs used for the rearing experiment were 460 nm (blue), 520 nm (green), 630 nm (red), and 450–680 nm (white). There was no notable difference in survival rate among fish in the four LED groups. However, the growth of juvenile PBT was lesser under red light compared to the green and white light wavelengths. These results suggest that PBT juveniles have low sensitivity to red light because the fish are rarely exposed to the red light wavelengths under natural ocean conditions. Thus, low sensitivity to red light negatively influenced the feeding behavior and growth of PBT juveniles.     

A Novel System for Spectral Analysis of Solar Radiation within a Mixed Beech-Spruce Stand

Abstract: A multi-sensor system is described, based on a 1024 channel diode array spectrometer, to measure spectral radiant flux density in the range of 380 nm to 850 nm, with a resolution of 0.8 nm in minimal 16 milliseconds integration time per sensor (noon, clear sky conditions). 264 space-integrating 4π sensors deployed in the canopies and 2 m above stand floor are sequentially connected to the spectrometer by means of 30-m long fibre optics. During low-level conditions (dawn, overcast sky) the system automatically lengthens the integration time of the spectrometer. About 3 sec per sensor, i.e., 13 min for the total of 264 sensors (worst case) are needed to collect spectral energy data, store them on hard disk and move the channel multiplexer to the next fibre optic position. The detection limit of quartz fibre sensors is 0.2 W/m²; precision and absolute error of radiant flux density are smaller than 3 % and 10 %, respectively.
The system, operating since 1999, is derived from a 20-sensor pilot system developed for PAR measurements (PMMA fibre sensor, 400nm to 700 nm).
Data achieved with the system serve to determine vertical profiles of wavelength dependent radiation extinction, with special respect to R/FR ratios and to develop a model of spectral radiation distribution in a mature forest stand, prerequisites for the computation of carbon gain of the stand and the evaluation of stand growth models.     

偏振干涉角度调制SPR传感技术研究及应用进展

针对一种新兴生物检测方法——表面等离子体波共振(SPR)技术,文中SPR传感系统采用偏振干涉和角度调制方案,使SPR传感灵敏度与光复反射系数的模和相位都相关,从而实现较大线形范围内的高灵敏测量。同时开展了该SPR传感系统在环保领域的应用研究,SPR共振信号可实时随甲烷含量线性改变,气体检测灵敏度达到1 070ppm,实验结果验证了这种SPR传感技术的检测性能并显示了其在环保监测领域的应用潜力。     

Integrated-optic biosensor by electro-optically modulated surface plasmon resonance

We present a new integrated-optic surface plasmon resonance (SPR) biosensor based on electro-optical modulation. The SPR characteristics for the analyte concentration detection can be electro-optically modulated by applying the voltage on the electrodes of the biosensor fabricated on lithium niobate, which is an excellent electro-optic material. Two measurement methods, electro-optically modulated SPR spectral measurement and electro-optically modulated SPR intensity measurement, are demonstrated and their measurands are the SPR wavelength and the output optical intensity, respectively. Human serum albumin is coated on the gold film surface of the proposed biosensor to detect the concentration of beta-blocker, which is a remedy for heart disease. As the applied voltage increases such that the effective index of guided mode rises, the SPR wavelength shifts toward the long wavelength side and the output optical intensity at the wavelength of 632.8 nm diminishes. The linear regression slope of the relation between the measurand and the applied voltage is dependent on the analyte concentration and can be used to determine the concentration variation. Experimental results measured by the electro-optically modulated SPR methods are compared with those measured by the conventional spectral and intensity methods, and the effects of waveguide width on the biosensor performance are discussed.