Sensitivity of surface plasmon resonance sensors based on metallic columnar thin films in the spectral and angular interrogations

Surface Plasmon Resonance (SPR) from metallic Columnar Thin Films (CTFs) of porosity as high as 0.5 was experimentally and theoretically investigated. The CTF layers were prepared by the Glancing Angle Deposition (GLAD) method. The SPR features were investigated in both the angular and the spectral modes. In the angular interrogation, increasing the porosity causes broadening to the dip width, shift to larger resonance angles, and increase of the sensitivity to analyte refractive index (RI) changes by about threefold compared with closed metal films. In the spectral interrogation, on the other hand, the resonance wavelengths are red-shifted for porous films; hence their spectral sensitivities are higher than those of closed films under the same experimental conditions. Nevertheless, the sensitivity behavior versus the resonance wavelength is similar to that of SPR sensors based on dense film layers. The shapes of the nanostructures constituting the CTF are described as ellipsoidal inclusions in which the effective permittivity dyadic of the composite material is calculated using the Bruggeman formalism with exact depolarization dyadics. The correlation between the sensitivity enhancement and the electromagnetic field intensity at the interface between the metallic film and the analyte was examined. Electromagnetic fields analyses were performed using the general 4 × 4 propagation matrices of general homogenous biaxial layers.     

Piezoelectric properties of microfabricated (K,Na)NbO3 thin films

A novel microfabrication method of lead-free piezoelectric sodium potassium niobate [(K,Na)NbO₃, KNN] thin films was proposed, and the piezoelectric characteristics of the KNN microactuators were evaluated. The KNN thin films were directly deposited on microfabricated Si microcantilevers. The transverse piezoelectric coefficient d₃₁ of the KNN films was calculated as −53.5 pm/V at 20 V_pp from the tip displacement of the microcantilevers. However, the tip displacement showed large electric-field dependence because of the extrinsic piezoelectric effect, and the intrinsic piezoelectric effect of the KNN microcantilevers was smaller than that of KNN on unprocessed thick substrates. In contrast, the extrinsic piezoelectric effect was almost independent of the microfabrication of the KNN films.     

Effects of heat treatment on oxidative gas adsorption for lead naphthalocyanine thin films

Thin films of Pb-naphthalocyanine (330 nm thick) prepared by vacuum sublimation were heat-treated at 250°C in air atmosphere for various periods. As estimated from the conductance versus temperature correlations, the adsorption of NO₂ and O₂ on the films at room temperature, which was only slight after the heat-treatment for 30 min, increased very remarkably with prolonging heat-treatment time up to 10 h. The 10 h treated film was found to exhibit completely reversible changes in conductance upon exposure to 5 ppm NO₂ in N₂ at 210°C. Visible light absorption spectra indicated a change from a largely amorphous state of the as-deposited film to a largely crystalline state of the film treated for 2 h. X-Ray diffraction analysis showed that the as-deposited film consisted of N-oriented particles, while the proportion of P-oriented particles increased with prolonging treatment time up to 10 h. The enhancement of gas adsorption was thus attributed to the reconstruction of films during the heat treatment. It appears that a reconstruction-assisted increase in porosity not only facilitates gas diffusion inside the film, but also increases the number of Pb-naphthalocyanine molecules accessible NO₂ or O₂.     

Interfacially polymerized metalloporphyrin thin films for colorimetric sensing of organic vapors

Interfacial polymerization chemistry has been employed to produce robustly structured supported films of sensor materials. Thin-film materials that incorporate several metalloporphyrins (M = Co(II), Cu(II) and Zn(II)) to use as colorimetric sensors for volatile organic compounds (VOCs) including alcohols, ketones and chlorocarbons have been synthesized. Upon exposure to these VOCs, the sensors’ absorption spectra undergo reversible changes that can be monitored to discriminate amongst analyte species. Sensor sensitivity and selectivity is affected by the metal identity.     

Ag-TiO2薄膜表面等离子体共振光谱特性研究

以拓宽Ag膜波长调制型表面等离子体共振(SPR)传感器的工作波长区间,将TiO2膜沉积于Ag膜表面制成Ag-TiO2复合薄膜.利用数值计算方法对不同厚度Ag膜和As-TiO2复合膜波长调制型SPR光谱特性进行仿真研究.仿真结果表明:共振吸收峰显著依赖于Ag膜厚度;当保持Ag-TiO2复合膜厚度60 nm时,复合薄膜SPR波长随TiO2厚度的增大而向长波长方向移动,发现红移;与60 nm厚Ag膜共振波长相比,12 nm厚TiO2与48 rnm厚Ag组成的复合膜共振波长红移超过200 nm.     

Enhanced chemosensing of ammonia based on the novel molecular semiconductor-doped insulator (MSDI) heterojunctions

A series of new molecular semiconductor-doped insulator (MSDI) heterojunctions as conductimetric transducers to NH₃ sensing were fabricated based on a novel semiconducting molecular material, an amphiphilic tris(phthalocyaninato) rare earth triple-decker complex, Eu₂[Pc(15C5)₄]₂[Pc(OC₁₀H₂₁)₈], quasi-Langmuir-Shäfer (QLS) film, as a top-layer, and vacuum-deposited and cast film of CuPc as well as copper tetra-tert-butyl phthalocyanine (CuTTBPc) QLS film as a sub-layer, named as MSDIs 1, 2 and 3, respectively. MSDIs 1-3 and respective sub-layers prepared from three different methods were characterized by X-ray diffraction, electronic absorption spectra and current-voltage (I-V) measurements. Depending on the sub-layer film-forming method used, α-phase CuPc film structure, β-phase CuPc crystallites and H-type aggregates of CuTTBPc have been obtained, respectively. An increasing sensitivity to NH₃ at varied concentrations in the range of 15-800 ppm, follows the order MSDI 2 < MSDI 3 < MSDI 1, revealing the effect of sub-layer film structures on sensing performance of the MSDIs. In particular, the time-dependent current plot of the MSDI 1, with α-phase CuPc film as a sub-layer, clearly shows an excellent separation of the different ammonia concentration levels and nearly complete reversibility and reproducibility even at room temperature, which is unique among the phthalocyanine-based ammonia sensors thus far reported in the literature. This provides a general method to improve sensor response of organic heterojunctions by controlling and tuning the film structure of sub-layer with appropriate fabrication techniques. On the other hand, the enhanced sensitivity, stability and reproducible response of the MSDI 1 heterostructure in comparison with the respective single-layer films have also been obtained. A judicious combination of materials and molecular architectures has led to enhanced sensing properties of the MSDI 1, in which control at the molecular level can be achieved.     

Effect of soft-hard segment structure on vapor responsiveness of polyurethane conducting composite thin films loaded with multi-walled carbon nanotubes

A series of multi-walled carbon nanotubes/polyurethane (MWNTs/PU) composite conducting dispersoids were prepared via an in situ coupling reaction among linear hydroxyl-terminated polymer diols, 1,6-hexamethylene diisocyanate (HDI) and various chain extenders. The composite conducting thin films were formed by spin-coating and depositing the dispersoids onto comb-like electrode substrates. The resulting structure and the dispersion quality of MWNTs in the dispersoids were examined by means of FTIR, XRD, TEM, SEM and UV-vis analyses. The response of the as-prepared films toward some volatile organic solvent vapors such as benzene, anhydrous ether, acetone and chloroform was evaluated. The experimental results indicated that the composite conducting films constructed by hydroxyl-terminated poly(butadiene-co-acrylonitrile), trimethylolpropane, and MWNTs-OH bear better vapor responsiveness. The dispersion behavior of MWNTs in the dispersoids, types of MWNTs and soft-hard segmental compositions are believed to be closely related with the sensing properties of the films. In particular, the chemical linkage of MWNT-OH with HDI in the PU matrix is expected to improve the dispersivity and further to enhance the sensing properties of the composite sensors. The vapor sensing properties well reveal that these materials have a possibility as a candidate of volatile organic solvent vapor sensors.     

Surface functionalization for self-referencing surface plasmon resonance (SPR) biosensors by multi-step self-assembly

Recently, a novel SPR sensor with on-chip referencing has been realized. In this sensor, one-half of the gold sensing surface is covered with a high refractive index overlayer of tantalum pentoxide (Ta₂O₅). When polychromatic beam illuminates the sensing surface, surface plasmon resonance in the areas with and without the overlayer occur at different wavelengths. Therefore, the reflected light exhibits two dips associated with SPRs in those two areas. When functionalized properly, one of the areas can be used as a specific sensing channel for detection of specific bio-interactions and the other can act as a reference channel for compensation for background refractive index fluctuations. In this paper we present a new functionalization approach for these mixed architecture chips. The gold side of the chip is functionalized with a mixed self-assembled monolayer of polyethylene oxide (PEO) and biotin terminated (BAT) thiols whereas the Ta₂O₅ side is coated with PEO terminated silanes. The PEO terminated thiols and silanes serve as a protein resistant background, while the biotin terminated thiols are used to bind streptavidin, which in turn immobilizes biotinylated antibodies. Hence, the gold side of the chip is used for the binding and detection of target analytes and the Ta₂O₅ side functions as a reference channel that monitors bulk refractive index changes and temperature drift. We have studied human chorionic gonadotropin (hCG) as a model system, currently detecting down to 5 ng/ml. In addition, we demonstrate the power of the on-chip reference channel for compensating for refractive index changes and eliminating false alarms.     

生物分子相互作用分析仪的自动控制系统设计

为达到光学表面等离子共振SPR传感的生物分子相互作用分析仪性能指标和功能要求,必须保证其自动控制系统的可靠性.针对时间相位调制SPR传感的特点,开发了一套自动控制系统.该系统利用CAN总线技术对仪器底层各个模块进行分布式控制,实现了按生物试验流程的自动控制.同时,采用等步距和等时间的离散加速度曲线法,分别控制盘库和波片机构的驱动电机,并采用数字PID控制流体池温度稳定和使CCD可靠制冷.试验结果表明,系统达到了分析仪的各项性能指标和功能要求.     

Surface plasmon resonance imaging (SPRi) as an alternative technique for rapid and quantitative screening of small molecules, useful in drug discovery

Surface plasmon resonance imaging (SPRi) is a label free technology for biomolecular interaction, which gives access to binding kinetic parameters from real time acquisition. It offers the possibility to test in a single run a large number of interactions, allowing rapid identification of the most suitable compounds toward a given biological entity. Until now, this technique has proven to be relevant for interaction between relatively large molecules (protein, antibodies, DNA) but has not been challenged yet for the screening of small molecules that can be of interested in the field of drug discovery. As a proof a principle, we have used SPRi to screen for interaction of several small molecules, referred to as G4-ligands on G-quadruplex DNA. This technology allowed to easy discrimination of the binding properties of four G4-ligands on quadruplex DNA models.     

Sensing properties of chemically sprayed TiO2 thin films using Ni, Ir, and Rh as catalysts

The purpose of this work is double, to analyze the influence of (i) the addition of different catalysts and (ii) the implementation of different procedures to introduce them in the titanium dioxide (TiO₂) thin films, in order to improve the film sensitivity for detecting oxygen. For reaching these objectives, TiO₂ thin films were deposited on alumina substrates by the ultrasonic spray pyrolysis (USP) technique employing titanium(IV) oxide acetylacetonate (TiO(acac)₂) as a titanium precursor, and pure methanol as a solvent. Iridium, nickel, and rhodium were the three catalysts used, which were incorporated by impregnation and USP onto the TiO₂ thin films surface. The electrical characterization, consisting in surface resistance measurements of the films, in a mixture of oxygen and zero-grade air, was performed using interdigitated gold electrodes contacted on the alumina substrates. From these, the film response or resistance change of the TiO₂ films was estimated. Single TiO₂ thin films measured at 400 °C displayed a response of the order of 0.02 and 0.18 to oxygen of 1000 and 10,000 ppm, respectively, whereas TiO₂ thin films using impregnated rhodium proved to have the highest response to O₂, with a value in the order of 2.5 to a concentration of 1000 ppm of O₂ diluted in zero-grade air at an optimal operating temperature of 250 °C.     

Sprayed CdIn2O4 thin films for liquefied petroleum gas (LPG) detection

Nanocrystalline cadmium indium oxide (CdIn₂O₄) thin films of different thicknesses were deposited by chemical spray pyrolysis technique and utilized as a liquefied petroleum gas (LPG) sensors. These CdIn₂O₄ films were characterized for their structural and morphological properties by means of X-ray diffraction (XRD) and scanning electron microscope (SEM), respectively. The dependence of the LPG response on the operating temperature, LPG concentration and CdIn₂O₄ film thickness were investigated. The results showed that the phase structure and the LPG sensing properties changes with the different thicknesses. The maximum LPG response of 46% at the operation temperature of 673 K was achieved for the CdIn₂O₄ film of thickness of 695 nm. The CdIn₂O₄ thin films exhibited good response and rapid response/recovery characteristics to LPG.     

Detection of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) by displacement of antibodies

A molecular layer with low non-specific binding enabling determination of low concentrations of 3,4-methylenedioxymethamphetamine (MDMA) by the displacement of antibodies has been developed. Antibody Fab′-fragments at various concentrations have been site-directly immobilised on gold and intercalated with a hydrophilic non-ionic polymer that reduces non-specific binding. Bovine serum albumin conjugated with MDMA and various concentrations of anti-MDMA antibodies were bound to the layer. The amount of conjugates and antibodies bound was dependent on the amount of Fab′-fragments in the layer. Antibodies were also bound to the conjugates physisorbed directly onto the gold surface and in mixtures with the polymer or with a lipoamide. A high displacement of antibodies was observed by surface plasmon resonance (SPR) on interaction of MDMA with the different layers in buffer solution. No displacement could, however, be observed in saliva with the pure conjugate layer because of a high non-specific binding of proteins. When the conjugates were coupled to the surface through the antibody Fab-fragment/polymer layer, MDMA concentrations as low as 0.02 ng mL⁻¹ (0.14 nM) could easily be detected in buffer. In diluted saliva the lowest limit of detection was 0.4 ng mL⁻¹ enabling determination of drugs from saliva with a cut-off concentration of 2 ng mL⁻¹. The molecular layer of antibody Fab′-fragments and polymer thus shows great potential for binding conjugates and antibodies that can be displaced on the interaction with very low concentrations of small-sized molecules. A low non-specific binding is guaranteed by the presence of the hydrophilic polymer.     

Surface plasmon resonance (SPR) sensors for the rapid, sensitive detection of the cellular response to osmotic stress

Cells regulate their volume in response to changes in osmolarity of both, their extracellular and intracellular environments. As stability of the cell volume is a compelling exigency for cellular integrity, techniques for a sensitive, time-resolved volume measurement of adherently grown mammalian cells attract considerable interest, especially in the field of cell physiology and biology. In this study we apply a surface plasmon resonance (SPR) based biosensor for the comparative analysis of the volume responses of two renal epithelial cell types to non-isotonic challenges. The on-line, label-free and non-invasive biosensor format shows distinct similarities and differences in the reaction kinetics of the two cell types. Furthermore regulatory volume responses to the osmotic stimuli as well as their inhibition by Gd³⁺ ions can be observed with a high time-resolution. Limit-of-detection measurements indicate the high sensitivity of the sensor capable of detecting cellular volume responses of adherently grown mammalian cells to osmotic stimuli well below a bioanalytical relevant value of 5 mOsm/kg.     

Signal enhancement of surface plasmon resonance-based immunoassays for the allergen detection

A surface plasmon resonance (SPR) biosensor was used to determine the recombinant group 1 house dust mite allergen (rDer f1) in both HBS-EP buffer and fetal bovine serum (FBS). The monoclonal antibody was immobilized onto the CM5 sensor chip surface using an amine coupling method. The procedures of antibody immobilization and the subsequent primary and enhanced immunoassay were monitored in real time. The sensitivity for rDer f1 detection was remarkably improved by using intact polyclonal antibody as signal amplifying agent. Using this signal enhanced SPR immunosensor, rDer f1 in HBS-EP buffer and FBS was detected at a concentration of 15.4 and 32.1 ng/ml, respectively. The result demonstrates that SPR biosensor is a simple and reliable method for allergen detection.     

Ochratoxin A immunoassay with surface plasmon resonance registration: Lowering limit of detection by the use of colloidal gold immunoconjugates

An immunoanalytical system was developed for the determination of ochratoxin A with the use of a surface plasmon resonance (SPR) sensor amplified by the anti-species antibody-colloidal gold particle (CGP) conjugate. The use of the binding of immune complexes to the CGP-anti-species antibody conjugate leads to the SPR signal amplification by a factor of more than 10 and results in the 60 pg/mL limit of detection of ochratoxin A with an assay time of 30 min. These characteristics are superior to those obtained both in the conventional enzyme immunoassay with the use of the same reagents and the SPR assay with unmodified antibodies and specific antibodies conjugated to colloidal gold.     

Approximation of trajectories of non-linear systems by iterates of systems with linear state dynamics

A modification of an iterative Picard process is proposed to approximate the output of a non-linear system by a concatenation of systems with linear state dynamics and non-linear outputs. A local uniform convergence result is given. The motivating example is a non-linear system that arises in surface plasmon resonance experiments to determine protein–protein interaction constants. We show with simulations that for this example the approximants converge not only locally but over the full time interval of interest in the application.     

Design and performances of immunoassay based on SPR biosensor with Au/Ag alloy nanocomposites

A novel method for detecting human IgG is reported, which is based on Au/Ag alloy nanocomposites for amplifying surface plasmon resonance response. Au/Ag alloy nanocomposites were characterized in detail by transmission electron microscopy (TEM), UV-vis absorption spectroscopy and X-ray photoelectron spectroscopy (XPS). Covalent immobilization of about 24 nm diameter of Au/Ag alloy nanocomposites on the Au film results in a large shift in resonance wavelength, which is due to the increase of the thickness of the sensing membrane, high dielectric constant of Au/Ag nanoparticles, and electromagnetic coupling between Au/Ag alloy nanocomposites and Au film. The SPR biosensor based on Au/Ag alloy nanocomposites exhibits a satisfactory response for human IgG in the concentration range of 0.15-40.00 μg mL⁻¹. While the biosensor based on Au nanoparticles shows a response in the concentration range of 0.30-20.00 μg mL⁻¹ and the biosensor based on Au film shows a response for human IgG in the concentration range of 1.25-20.00 μg mL⁻¹.     

Detection of sulphamethazine with an optical biosensor and anti-idiotypic antibodies

We describe the development of an assay for the detection of sulphamethazine in animal urine with a surface plasmon resonance (SPR) biosensor. In order to obtain a general assay that can easily be transferred to other veterinary drug residues, a monoclonal antibody against sulphamethazine and a corresponding anti-idiotypic antibody were used. The assay had a lower detection limit of 5 μg/l which is well below the maximum residue limits (MRL) of 100 μg/l and can be used for screening purposes in animal urine. The binding reaction between the antibodies as occurring during sensor application was characterized with respect to avidity and kinetic properties.     

Label-free sensitive optical detection of polychlorinated biphenyl (PCB) in an aqueous solution based on surface plasmon resonance measurements

A label-free, ultrasensitive method for the optical detection of polychlorinated biphenyl (PCB) is described. The detection mechanism for PCB is based on PCB-induced conformational changes of immobilized Cytochrome c (Cyt c) on an Au thin film altering the local dielectric function of the supported Cyt c, which can be detectable by surface plasmon resonance (SPR) spectroscopy. Results showed that the confirmations of Cyt c changed in a very sensitive way depend on PCB concentrations as confirmed by circular dichroism (CD) measurements and atomic force microscope (AFM) observations. Based on the results, we investigated the PCB-sensitive detection performance of the self-assembling Cyt c monolayer surface by SPR. On exposure to PCB, the reflectance R at the SPR angle of the supported Cyt c showed a sensitive and systematic increase with increasing PCB concentrations. The limit of detection was as low as 0.1 ppb and the responses completed within 10 min.