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1.
Functionalization of a gold surface is usually accomplished by covalent binding via self-assembled monolayers (SAMs) on the gold surface, followed by attachment of flexible polymeric linker layers such as dextran hydrogels. However, these techniques require multiple steps and also have nonspecific interactions and steric problems. In this study, a self-assembled carboxylated terthiophene monolayer was formed onto a gold surface to create a sensitive and stable surface plasmon resonance (SPR) biosensing system. Compared with a commercial carboxymethyl dextran chip (CM5), the terthiophene SAM surface provided more than six times more antibody-binding signals and nearly three times the SPR assay sensitivity for progesterone (P4).  相似文献   

2.
We enhanced the sensitivity of surface plasmon resonance biosensor by the conversion of the real-time direct binding immunoassay into the sandwich immunoassay, in which colloidal gold particles coated with anti-mouse IgG was used. By the immobilization of anti-mouse IgG onto the carboxymethyl dextran surface of thin gold film, the direct binding of analyte (mouse IgG) onto the sensor chip, and the injection of colloidal gold particles coated with antimouse IgG, about 100 times of sensitivity enhancement was obtained. This result suggests that nanoparticles, which has a high refractive index, homogeneous ultrafine structure and capability of size control, would be applicable for the detection of very small quantity of biomaterial.  相似文献   

3.
Surface plasmons resonance (SPR) architectures based on grating coupler/disperser combination is an attractive alternative for spectral-based biochemical sensing. In this paper, we investigate theoretically and experimentally a new concept where the plasmon coupling occurs through a thin film grating and sensing occurs via the first evanescent diffraction order in transmitive mode. The surface plasmon wave excitation induces a peak in the wavelength as well as in the angular spectra of the detected first transmitted diffraction order. Accordingly, a change in SPR spectrum of the detected diffraction order can be used to quantify the amount of the target molecules immobilized on the sensor surface, and therefore, the concentration of these molecules in the analyte solution. The developed sensor architecture is dedicated to droplet biochemical sensing and appears to be especially suitable for biosensor integration and miniaturization. The presented sensor concept is perfectly suited for mass production of low-cost and reproducible SPR sensor chip for biochemical analysis. The implemented setup gives access to multichannel biosensing with the potential for efficient internal referencing essential to achieve sufficiently high reproducibility and accuracy of the measurements.  相似文献   

4.
A novel sensing method based on surface plasmon resonance (SPR) was developed for the highly sensitive quantification of low molecular weight (LMW) analytes (colloidal Au replacement assay). Gold nanoparticles (diameter = 20 nm) functionalized with lactosyl-poly(ethylene glycol) (PEG) were prepared and were specifically adsorbed onto a Ricinus communis agglutinin (RCA120)-immobilized SPR sensor chip surface. Subsequent injection of free d-galactose elicited the elution of the preadsorbed lactosyl-PEGylated gold nanoparticles in a manner proportional to the galactose concentration, achieving a substantial and quantitative analysis over a wide range of galactose concentrations (0.1-50 ppm). This method of d-galactose sensing through the substituted elution of preadsorbed nanoparticles from the sensor chip surface would be applicable for the highly sensitive SPR quantification of various LMW analytes, which are known to be difficult to detect by the conventional SPR sensing regime.  相似文献   

5.
An immobilisation procedure based on the direct coupling of thiol-derivatised oligonucleotide probes to bare gold sensor surfaces has been used for DNA sensing applications. The instrumentation used relies on surface plasmon resonance (SPR) transduction; in particular the commercially available instruments BIACORE X and SPREETA, have been employed in this study. The performances of the SPR-based DNA sensors resulting from direct coupling of thiol-derivatised DNA probes onto gold chips, have been studied in terms of the main analytical parameters, i.e. selectivity, sensitivity, reproducibility, analysis time, etc. A comparison between the thiol-derivatised immobilisation approach and a reference immobilisation method, based on the coupling of biotinylated oligonucleotide probes onto a streptavidin coated dextran sensor surface, using synthetic complementary oligonucleotides has been discussed. Finally, a denaturation method to obtain ssDNA ready for hybridisation analysis has been applied to polymerase chain reaction (PCR) amplified samples, for the detection of genetically modified organisms (GMOs).  相似文献   

6.
We developed a novel technique for increasing the sensitivity of transmission surface plasmon resonance (T-SPR) signals. T-SPR spectroscopy was performed by irradiating, with white light, a gold grating substrate whose surface was nanostructured by growing gold nanoparticles (AuNPs). AuNPs were grown directly on the substrate surface by alcohol reduction and their growth was observed at various stages by UV–visible spectroscopy and standard Kretschmann-type SPR spectroscopy. For comparison, normal gold film with smooth surface was examined under similar condition. The T-SPR results show a possibility of hybrid excitation of both localized and propagating surface plasmon. Significantly, T-SPR spectra of the gold grating substrate obtained during AuNP growth show stronger and narrower peaks in the range 650–800 nm. The maximum T-SPR excitation was at an incident angle of 35°. A layer-by-layer system of 5,10,15,20-tetrakis (1-methyl-4-pyridinio)porphyrin tetra(p-toluenesulfonate) molecules and sodium copper chlorophyllin molecules was used to verify the enhancement of the developed system. We believe that the AuNPs/Au grating for T-SPR devices will provide enhanced signals for detecting nanometer order materials and for high-sensitive sensor applications.  相似文献   

7.
Plasmonics - In this study, we achieved an enhancement of the transmission surface plasmon resonance (T-SPR) intensity by depositing silver nanoparticles (AgNPs) onto a gold grating substrate. The...  相似文献   

8.
We demonstrate directional enhanced fluorescence emission from fluorophores located above gold wire gratings. In contrast to previous studies on corrugated films, efficient coupling was recorded for multiple plasmon modes associated with both the active and substrate side of the wires. This difference is likely due to the subtle differences in how light interacts with corrugated films versus metal films with periodic subwavelength slots. For corrugated films, coupling between modes on opposite sides of the grating are out of phase, and therefore plasmon modes on the opposite side of the grating are only weakly excited. For wire gratings, transmission and reflection features have been modeled well with a dynamical diffraction model that includes surface plasmons, which allows for efficient coupling to surface plasmon modes on both sides of the grating. We also compared the two mechanisms for fluorescent enhancement, namely the intense electromagnetic field associated with surface plasmons and excited fluorophores radiating via surface plasmon modes. We found the latter mechanism clearly dominant.  相似文献   

9.
Grating-coupled surface plasmon resonance (GCSPR) is a method for the accurate assessment of analyte in a multiplexed format using small amounts of sample. In GCSPR, the analyte is flowed across specific receptors (e.g. antibodies or other proteins) that have been immobilized on a sensor chip. The chip surface is illuminated with p-polarized light that couples to the gold surface's electrons to form a surface plasmon. At a specific angle of incidence, the GCSPR angle, the maximum amount of coupling occurs, thus reducing the intensity of reflected light. Shifts in the GCSPR angle can be correlated with refractive index increases following analyte capture by chip-bound receptors. Because regions of the chip can be independently analyzed, this system can assess 400 interactions between analyte and receptor on a single chip. We have used this label-free system to assess a number of molecules of immunological interest. GCSPR can simultaneously detect an array of cytokines and other proteins using the same chip. Moreover, GCSPR is also compatible with assessments of antigen expression by intact cells, detecting cellular apoptosis and identifying T cells and B cells. This technology represents a powerful new approach to the analysis of cells and molecular constituents of biological samples.  相似文献   

10.
A small array of subwavelength apertures patterned in a gold film on glass was characterized for use as a biosensor. It is widely believed that such arrays allow the resonance of photons with surface plasmons in the metallic film. Surface plasmon methods (and other evanescent wave methods) are extremely well suited for the measure of real time biospecific interactions. An extremely high sensitivity of 88,000%/refractive index unit was measured on an array with theoretical active area of .09 microm2. The formation of a biological monolayer was monitored. Both sensitivity and resolution were determined through measurement. The measured resolution, for a sensor with an active area of less than 1.5 microm2, is 9.4 x 10(-8) refractive index units which leads to a calculated sensitivity of 3.45E6%/refractive index unit. These values far exceed theoretical and calculated values of other grating coupled surface plasmon resonance (SPR) detectors and prism based SPR detectors. Because the active sensing area can be quite small (.025 microm2) single molecule studies are possible as well as massive multiplexing on a single chip format.  相似文献   

11.
We report a simple 1D grating device fabrication on ~50 nm gold (Au) film deposited on glass, which is employed as a high performance refractive index (RI) sensor by exploiting the surface plasmon polaritons (SPP) excited by the grating device along the Au/analyte interface. A finite element analysis (FEA) method is employed to maximize the sensitivity of the sensor for a fixed period and thickness of a gold film and its close correspondence with experiment has given the insight for high sensitivity and enhanced transmission. Significantly, in the context of economic design and performance, it is shown that an optimally designed and fabricated 1D grating can be as sensitive as 524 nm/RIU (linearity RI?=?1.33303 to 1.47399), which is remarkably higher than existing reports operating in a similar wavelength region.  相似文献   

12.
13.
Effective surface immobilization is a prerequisite for numerous carbohydrate-related studies including carbohydrate-biomolecule interactions. In the present work, we report a simple and rapid modification technique for diverse carbohydrate types in which direct oriented immobilization onto a gold surface is accomplished by coupling the amine group of a thiol group-bearing aminophenyl disulfide as a new coupling reagent with an aldehyde group of the terminal reducing sugar in the carbohydrate. To demonstrate the generality of this proposed reductive amination method, we examined its use for three types of carbohydrates: glucose (monosaccharide), lactose (disaccharide), and GM1 pentasaccharide. Through successful mass identifications of the modified carbohydrates, direct binding assays on gold surface using surface plasmon resonance and electrochemical methods, and a terminal galactose-binding lectin assay using atomic force microscopy, we confirmed several advantages including direct and rapid one-step immobilization onto a gold surface and exposure of functional carbohydrate moieties through oriented modification of the terminal reducing sugar. Therefore, this facile modification and immobilization method can be successfully used for diverse biomimetic studies of carbohydrates, including carbohydrate-biomolecule interactions and carbohydrate sensor or array development for diagnosis and screening.  相似文献   

14.
A surface plasmon resonance (SPR) imaging system was constructed and used to detect the hexahistidine-ubiquitin-tagged human parathyroid hormone fragment (His6-Ub-hPTHF(1–34)) expressed inEscherichia coli. The hexahistidine-specific antibody was immobilized on a thin gold film coated with ProLinkerTM B, a novel calixcrown derivative with a bifunctional coupling property that permits efficient immobilization of capture proteins on solid matrices. The soluble and insoluble fractions of anE. coli cell lysate were spotted onto the antibody-coated gold chip, which was then washed with buffer (pH 7.4) solution and dried. SPR imaging measurements were carried out to detect the expressed His6-Ub-hPTHF (1–34). There was no discernible protein image in the uninduced cell lysate, indicating that non-specific binding of contaminant proteins did not occur on the gold chip surface. It is expected that the approach used here to detect affinity-tagged recombinant proteins using an SPR imaging technique could be used as a powerful tool for the analyses of a number of proteins in a high-throughput mode.  相似文献   

15.
Surface plasmon resonance biosensors depend on modified gold surfaces to allow immobilization of proteins or peptides for interaction analysis. We investigated sensor chip surfaces that differ in the geometry of the immobilization matrix: two contain a three-dimensional coupling matrix and two have a surface with immobilization sites on a two-dimensional plane. Properties of sensor chips were compared by studying the interaction of calmodulin with a peptide representing the calmodulin-binding site of nitric oxide synthase I. Apparent K(D) values were determined by three different procedures in order to apply tests for self-consistency. At low surface densities (5-8 fmol/mm(2)) on three of the four tested surfaces, estimated K(D) values were within one order of magnitude and similar to the value found in solution (K(D) = 1-3 nM). When immobilization densities were increased by one to two orders of magnitude, apparent association rate constants were less distorted on a flat carboxymethylated surface than on dextran-coated sensor chips.  相似文献   

16.
A novel plasmonic structure based on an anticrossing bandgap prism coupling technique is proposed. The study has been carried out using photonic crystals based on diffraction gratings (bounded by dielectrics with identical dielectric functions) together with a high refractive index prism to couple the long-range surface plasmon polaritons to photons. We analyse the structure and demonstrate the ability for tuning the propagation constants of plasmon modes by changing the thickness of the gold grating. The comparison to non-bandgap techniques is studied, and the influence of the plasmonic configuration on the plasmon propagation constant is discussed as well. Experimental measurements were also carried out to confirm the validity of our model.  相似文献   

17.
We propose a surface plasmon resonance (SPR)-based refractive index sensor using gold-alumina grating over aluminum film for biosensing. Conventional SPR sensor based on gold grating exhibits broader SPR dips whereas that based on aluminum grating exhibits narrow reflection dip. A narrow reflection dip is desirable as it provides good resolution and improves the accuracy of measurement. Aluminum is less stable and generally is not preferred for an SPR-based sensor. It is more prone to being oxidized, which reduces the sensitivity and increases the width of the reflection dip of the sensor. While gold cannot provide narrow SPR reflection dips, but is used as an SPR active metal due to its more chemical stability. In order to improve the accuracy of gold grating-based sensor while taking care of oxidation problem of aluminum, in this paper, we propose a gold grating over aluminum film for SPR-based sensor and show that this configuration improves the sensitivity and the detection accuracy of the conventional sensor. Moreover, the oxidation problem is reduced to some extent as a part of aluminum is covered with gold. In order to completely avoid the oxidation of aluminum, we further propose to cover the exposed part of the aluminum with alumina and show that this configuration further improves the accuracy by reducing the width of the SPR reflection dip without affecting the sensitivity significantly. Numerical simulations show that sensitivity of proposed sensor is 270.33°/RIU with quality factor of more than 267.65 RIU?1.  相似文献   

18.
Grating-coupled surface plasmon resonance imaging (GCSPRI) is a method for the accurate assessment of both cell phenotype and function. In GCSPRI, cells and/or proteins of interest are flowed across antibodies immobilized on a gold-coated sensor chip. The surface of the chip is illuminated with monochromatic light that couples with surface plasmons in the gold. At a specific angle of incidence, the GCSPR angle, the maximum amount of coupling occurs. Shifts in the GCSPR angle can be correlated with refractive index changes following cell or analyte capture by the immobilized antibodies. In addition, GCSPRI can image the cells as they are being captured. GCSPRI's multiplexed format allows for the parallel assessment of up to 400 individual antibody regions. In this paper, we demonstrate GCSPRI's ability to identify cells and proteins of interest and compare results to a traditional flow cytometry system. This technology represents a fast and powerful method for the simultaneous assessment of cell phenotype and function.  相似文献   

19.
A molecularly imprinted polymer (MIP) film for domoic acid (DA) was synthesised by direct photo-grafting onto a gold chip suitable for a surface plasmon resonance (SPR) based bioanalytical instrument system, the BIAcore 3000. The gold surface was first functionalised with a self-assembled monolayer of 2-mercaptoethylamine and subsequent carbodiimide chemistry was performed for covalent attachment of the photoinitiator, 4,4'-azobis(cyanovaleric acid). This ensured that the formation of the MIP thin film, comprising 2-(diethylamino) ethyl methacrylate as functional monomer and ethylene glycol dimethacrylate as cross-linker, occurred only at the surface level. Optimisation and control over the grafting procedure were achieved using contact angle measurements and atomic force microscope (AFM) imaging. The surface grafting resulted in the formation of thin and homogeneous MIP film with thickness of 40 nm. A competitive binding assay was performed with free DA and its conjugate with horseradish peroxidase, which was used as a refractive label. The sensor was evaluated for its sensitivity, cross-reactivity, and robustness by using a BIAcore 3000. Likewise, monoclonal antibodies acting as natural receptors for the toxin were studied with the same BIAcore system. Results of a comparison between the artificial and natural receptors are reported. In contrast to monoclonal antibodies, the regeneration of MIP chip did not affect its recognition properties and continuous measurement was possible over a period of at least 2 months.  相似文献   

20.
In this report, we have investigated enhanced surface plasmon resonance (SPR) detection of DNA hybridization using gold core - silica shell nanoparticles in localized plasmonic fields. The plasmonic fields were localized by periodic linear gratings. Experimental results measured for hybridization of 24-mer single-stranded DNA oligomers suggest that core-shell nanoparticles (CSNPs) on gratings of 400 nm period provide enhanced optical signatures by 36 times over conventional thin film-based SPR detection. CSNP-mediated DNA hybridization produced 3 times larger angular shift compared to gold nanoparticles of the same core size. We have also analyzed the effect of structural variation. The enhancement using CSNPs was associated with increased surface area and index contrast that is combined by improved plasmon coupling with localized fields on gratings. The combined approach for conjugated measurement of a biomolecular interaction on grating structures is expected to lower the limit of detection to the order of a few tens of fg/mm(2).  相似文献   

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