Optimization of the Field Enhancement and Spectral Bandwidth of Single and Coupled Bimetal Core–Shell Nanoparticles for Few-Cycle Laser Applications

We have theoretically studied and optimized the field enhancement and temporal response of single and coupled bimetal Ag/Au core–shell nanoparticles (NPs) with a diameter of 160 nm and compared the results to pure Ag and Au NPs. Very high-field enhancements with an amplitude reaching 100 (with respect to the laser field centered at 800 nm) are found at the center of a 2-nm gap between Ag/Au core–shell dimers. We have explored the excitation of the bimetal core–shell particles by Fourier transform-limited few-cycle optical pulses and identified conditions for an ultrafast plasmonic decay on the order of the excitation pulse duration. The high-field enhancement and ultrafast decay makes bimetal core–shell particles interesting candidates for applications such as the generation of ultrashort extreme ultraviolet radiation pulses via nanoplasmonic field enhancement. Moreover, in first experimental studies, we synthesized small bimetal Ag/Au core–shell NPs and compared their optical response with pure Au and Ag NPs and numerical results.     

Preparation of Ag/Au bimetallic nanostructures and their application in surface‐enhanced fluorescence

An effective substrate for surface‐enhanced fluorescence, which consists of cluster Ag/Au bimetallic nanostructures on a copper surface, was synthesized via a multi‐stage galvanic replacement reaction of a Ag cluster in a chlorauric acid (HAuCl₄) solution at room temperature. The fabricated silver/gold bimetallic cluster were found to yield large surface‐enhanced fluorescence (SEF) enhancement factors for rhodamine 6G probe molecules deposited on the substrate, and also the fluorescence efficiency is critically dependent on the period of nanostructure growth. With the help of proper control reaction conditions, such as the reaction time, and concentration of reaction solutions, the maximum fluorescence enhanced effect was obtained. Therefore, the bimetallic nanostructure substrate also can be adapted to studies in SEF, which will expand the application of SEF. Copyright © 2015 John Wiley & Sons, Ltd.     

Sensitivity enhancement of surface plasmon resonance biosensor with colloidal gold

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.     

Directional surface plasmon-coupled emission: application for an immunoassay in whole blood

We present a new approach for performing fluorescence immunoassay in whole blood using fluorescently labeled anti-rabbit immunoglobulin G (IgG) on a silver surface. This approach, which is based on surface plasmon-coupled emission (SPCE), provides increased sensitivity and substantial background reduction due to exclusive selection of the signal from the fluorophores located near a bioaffinity surface. This article describes the effect of an optically dense sample matrix, namely human whole blood and serum, on the intensity of the SPCE. An antigen (rabbit IgG) was adsorbed to a slide covered with a thin silver metal layer, and the SPCE signal from the fluorophore-labeled anti-rabbit antibody, binding to the immobilized antigen, was detected. The effect of the sample matrix (buffer, human serum, or human whole blood) on the end-point immunoassay SPCE signal was studied. It was demonstrated that the kinetics of binding could be monitored directly in whole blood or serum. The results showed that human serum and human whole blood attenuate the SPCE end-point signal and the immunoassay kinetic signal only approximately two- and threefold, respectively, as compared with buffer, resulting in signals that are easily detectable even in whole blood. The high optical absorption of the hemoglobin can be tolerated because only fluorophores within a couple of hundred nanometers from the metallic film contribute to SPCE. Excited fluorophores outside the 200-nm layer do not contribute to SPCE, and their free space emission is not transmitted through the opaque metallic film into the glass substrate. We believe that SPCE has the potential of becoming a powerful approach for performing immunoassays based on surface-bound analytes or antibodies for many biomarkers directly in dense samples such as whole blood with no need for washing steps.     

The Effect of Bumpy Structure on Optical Properties of Bimetallic Nanoshells

In this paper, a sensitive bumpy bimetallic nanoshell for detection of thyroid cancer market (Thyroglobulin, Tg) and bovine serum albumin (BSA) proteins is reported. The physical origin of plasmonic properties of bimetal nanoshells is described by plasmon hybridization theory which indicates three intense and clearly separated plasmon modes. The electric field intensity enhancement of the bumpy bimetal nanoshell increases by ~559 %, at the surface of the bump in comparison with a smooth shell. The presence of bumpy structure on the nanoshell surface provides a high enhancement of the resulting Raman signal through an electromagnetic field of the order of 10⁷ which leads to an increase in sensitivity detection of Tg and BSA proteins. In addition, a refractive index (RI) sensitivity of 332.54 nm/RIU is achieved for this bumpy bimetallic nanoshell.     

Effect of dielectric spacer thickness on signal intensity of surface plasmon field-enhanced fluorescence spectroscopy

Surface plasmon field-enhanced fluorescence spectroscopy (SPFS) combines enhanced field platform and fluorescence detection. Its advantages are the strong intensity of the electromagnetic field and the high signal/noise (S/N) ratio due to the localized evanescent field at the water/metal interface. However, the energy transfer from the fluorophore to the metal surface diminishes the fluorescence intensity, and this reduces the sensitivity. In this study, we tested whether polystyrene (PSt) could act as a dielectric layer to suppress the energy transfer from the fluorophore to the metal surface. We hypothesized that this would improve the sensitivity of SPFS-based immunoassays. We used α-fetoprotein (AFP) as a model tumor biomarker in the sandwich-type immunoassay. We determined the relationship between fluorescent signal intensity and PSt layer thickness and compared this to theoretical predictions. We found that the fluorescence signal increased by optimally controlling the thickness of the PSt layer. Our results indicated that the SPFS-based immunoassay is a promising clinical diagnostic tool for quantitatively determining the concentrations of low-level biomarkers in blood samples.     

Cross-talk-free multiplexed immunoassay using a disposable electrochemiluminescent immunosensor array coupled with a non-array detector

A disposable electrochemiluminescent (ECL) immunosensor array was fabricated on a screen-printed carbon electrode (SPCE) substrate to perform multiplexed immunoassay (MIA) for the first time. The SPCE substrate was composed of an array of four carbon working electrodes, one common Ag/AgCl reference electrode, and one common carbon counter electrode. The immunosensor array was constructed by site-selectively immobilizing multiple antigens on different working electrodes of the SPCE substrate. With a competitive immunoassay format, the immobilized antigens competed with antigens in the sample to capture their corresponding tri(2,2'-bipyridyl)ruthenium(II)-labeled antibodies. The ECL signals from the immunosensors in this array were sequentially detected by a photomultiplier with the aid of a homemade single-pore-four-throw switch. Due to the ECL readout mechanism and the sequential detection mode, it could avoid the cross-talk between the adjacent immunosensors, which was common in other reported immunosensor array. Human, rabbit and mouse immunoglobulin Gs were near-simultaneously assayed as the model analytes. The linear ranges for them were 10-400, 20-400, and 20-400 ng/mL, with detection limits of 2.9, 6.1 and 6.5 ng/mL (S/N=3), respectively. This novel ECL strategy based on immunosensor array coupled with non-array detector provided a simple, sensitive, low-cost and time-saving approach for MIA. It showed great application potential in point-of-care test and field analysis of bio-agents, with mass production potential and high throughput.     

The Effect of Ag Nanoparticles on Surface-Enhanced Luminescence from Au Nanovoid Arrays

Studies comparing the effect of adding two different nanoparticle compositions on the plasmonic properties of Au nanovoid arrays were undertaken. Surface-enhanced resonance luminescence and surface-enhanced resonance Raman studies comparing dispersed Ag nanoparticles and Ag nanoparticle aggregates on gold nanovoid arrays were undertaken. These studies showed that using Ag nanoparticle aggregates increased both luminescence and Raman efficiency relative to when dispersed nanoparticles were used; in addition, these studies also showed that adding dispersed Ag nanoparticles supported a more reproducible enhancement in luminescence and Raman across the substrate compared to using Ag nanoparticle aggregates. Finite element analysis simulations indicated that surface plasmon polariton distribution in the sample was affected by the presence of the Ag nanoparticles on the Au nanovoid array.     

Use of surface plasmon-coupled emission to measure DNA hybridization

The authors describe a new approach to measuring DNA hybridization based on surface plasmon-coupled emission (SPCE). SPCE is the resonance coupling of excited fluorophores with electron motions in thin metal films, resulting in efficient transfer of energy through the film and radiation into the glass substrate. The authors evaluated the use of SPCE for detection of DNA hybridization. An unlabeled capture biotinylated oligonucleotide was attached near the surface of a thin (50 nm) silver film using streptavidin. The authors then measured the emission intensity of single-stranded Cy5-labeled DNA upon binding to a complementary oligomer attached to a silver film. Hybridization could be detected by an increase in SPCE, which appeared as light radiated into the substrate at a sharply defined angle near 73 degrees from the normal. The largest signals were observed when the excitation angle of incidence equaled the surface plasmon wavelength, but directional emission was also observed without excitation by the surface plasmon evanescent field. The increased intensity is due to proximity to the metal surface, so that hybridization can be detected without a change in the quantum yield of the fluorophore. These results indicate that SPCE can provide highly sensitive real-time measurement of DNA hybridization.     

Sequential use of the PAP and immunogold staining methods for the light microscopical double staining of tissue antigens: Its application to the study of regulatory peptides in the gut

Double immunoperoxidase staining using different couplers can give various combinations of colours on a single tissue section to achieve a comparable picture of different antigens. However, the colour combinations achieved to date are not entirely satisfactory.A double immunostaining procedure is introduced here, combining the peroxidase anti-peroxidase (PAP) and immunogold staining (IGS) methods. The IGS method is a new, simple, sensitive and reliable approach to immunostaining at the light microscopic level. It was carried out in three ways. Firstly, a two-step method was used in which the second layer was goat anti-rabbit IgG adsorbed onto gold particles (GAR/Au20). Secondly, a three-step method was employed where the second layer was unlabelled goat anti-rabbit IgG and the third layer was a rabbit antibody to peroxidase adsorbed onto the gold particles (RAP/Au20) and acting as a gold-labelled IgG antigen. The third method combined the first two methods using GAR/Au20 as the second layer and RAP/Au20 as the third layer which increased the amount of bound gold and enhanced the red colour, providing a better picture.The use of gold-labelled antibodies in double immunostaining has great potential value for many studies including that of the diffuse neuroendocrine system of the gut.     

Dopamine-Induced Growth of Au and Ag Nanoparticles on ITO Substrate and Their Application in PCPDTBT-Based Polymer Solar Cell

The direct attachment and growth of gold or silver nanoparticles (NPs) on indium tin oxide (ITO) surfaces was demonstrated using a simple and inexpensive successive ionic layer adsorption and reaction (SILAR) method by chemical reduction of the precursor metal salts with dopamine aqueous solution. Ag NPs on ITO substrate were approximately spherical with an average particle size of about 57 nm, but had a wide particle size distribution. Compared with Ag NPs, under the same 10 SILAR cycles, Au NPs have higher density packing and smaller average particle size of about 36 nm. XRD characterization and surface chemistry analysis confirmed the formation of Ag and Au NPs on ITO substrate with small amounts of dopamine-quinone adsorbed on the surface of them. Although Au NPs showed characteristic plasmon absorption, this did not result in performance enhancement in solar cell with the structure of ITO/ZnO/PCPDTBT:[6,6]-phenyl C₇₁/MoO₃/Ag because of the energy level mismatch between ZnO and dopamine molecules adsorbed on the surface of metal NPs.     

Modification of silicon nanoparticle surface with gold or silver attenuates its biocompatibility in vitro

The effects of crystalline silicon (Si) nanoparticles covered with gold and silver on the viability and state of cellular organelles of cultured human peripheral blood lymphocytes were investigated. After interaction with Si nanoparticles covered with gold (Si/Au) or with silver (Si/Ag), the reactive oxygen species in cells increased, but their viability was not decreased. The Si/Au nanoparticles decreased functional activity of lysosomes and mitochondria, while Si/Ag decreased the functional activity only of mitochondria. It is concluded that modification of a surface with gold or silver results in a reduction of biocompatibility of crystalline silicon nanoparticles.     

Microfluidic systems integrated with two-dimensional surface plasmon resonance phase imaging systems for microarray immunoassay

This study reports a microfluidic chip integrated with an arrayed immunoassay for surface plasmon resonance (SPR) phase imaging of specific bio-samples. The SPR phase imaging system uses a surface-sensitive optical technique to detect two-dimensional (2D) spatial phase variation caused by rabbit immunoglobulin G (IgG) adsorbed on an anti-rabbit IgG film. The microfluidic chip was fabricated by using micro-electro-mechanical-systems (MEMS) technology on glass and polydimethylsiloxane (PDMS) substrates to facilitate well-controlled and reproducible sample delivery and detection. Since SPR detection is very sensitive to temperature variation, a micromachine-based temperature control module comprising micro-heaters and temperature sensors was used to maintain a uniform temperature distribution inside the arrayed detection area with a variation of less than 0.3 degrees C. A self-assembled monolayer (SAM) technique was used to pattern the surface chemistry on a gold layer to immobilize anti-rabbit IgG on the modified substrates. The microfluidic chip is capable of transporting a precise amount of IgG solution by using micropumps/valves to the arrayed detection area such that highly sensitive, highly specific bio-sensing can be achieved. The developed microfluidic chips, which employed SPR phase imaging for immunoassay analysis, could successfully detect the interaction of anti-rabbit IgG and IgG. The interactions between immobilized anti-rabbit IgG and IgG with various concentrations have been measured. The detection limit is experimentally found to be 1 x 10(-4)mg/ml (0.67 nM). The specificity of the arrayed immunoassay was also explored. Experimental data show that only the rabbit IgG can be detected and the porcine IgG cannot be adsorbed. The developed microfluidic system is promising for various applications including medical diagnostics, microarray detection and observing protein-protein interactions.     

Application of surface plasmon coupled emission to study of muscle

Muscle contraction results from interactions between actin and myosin cross-bridges. Dynamics of this interaction may be quite different in contracting muscle than in vitro because of the molecular crowding. In addition, each cross-bridge of contracting muscle is in a different stage of its mechanochemical cycle, and so temporal measurements are time averages. To avoid complications related to crowding and averaging, it is necessary to follow time behavior of a single cross-bridge in muscle. To be able to do so, it is necessary to collect data from an extremely small volume (an attoliter, 10(-18) liter). We report here on a novel microscopic application of surface plasmon-coupled emission (SPCE), which provides such a volume in a live sample. Muscle is fluorescently labeled and placed on a coverslip coated with a thin layer of noble metal. The laser beam is incident at a surface plasmon resonance (SPR) angle, at which it penetrates the metal layer and illuminates muscle by evanescent wave. The volume from which fluorescence emanates is a product of two near-field factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. The fluorescence is quenched at the metal interface (up to approximately 10 nm), which further limits the thickness of the fluorescent volume to approximately 50 nm. The fluorescence is detected through a confocal aperture, which limits the lateral dimensions of the detection volume to approximately 200 nm. The resulting volume is approximately 2 x 10(-18) liter. The method is particularly sensitive to rotational motions because of the strong dependence of the plasmon coupling on the orientation of excited transition dipole. We show that by using a high-numerical-aperture objective (1.65) and high-refractive-index coverslips coated with gold, it is possible to follow rotational motion of 12 actin molecules in muscle with millisecond time resolution.     

A chemiluminescent metalloimmunoassay based on silver deposition on colloidal gold labels

A sensitive chemiluminescent (CL) immunoassay of human immunoglobulin (IgG) which combined the inherent high sensitivity of CL analysis with the dramatic signal amplification of silver precipitation on colloidal gold tags was developed. First, the sandwich-type complex was formed in this protocol by the primary antibody immobilized on the polystyrene wells, the analyte in the sample, and the secondary antibody labeled with colloidal gold. Second, the colloidal gold was treated by an Ag(+) reduction solution, which resulted in the catalytic precipitation of silver on the surface of colloidal gold. Third, a large number of Ag(+) were oxidatively released in HNO(3) solution from the silver metal anchored on the sandwich-type complexes and then the human IgG was indirectly determined by a sensitive combined CL reaction of Ag(+)-K(2)S(2)O(8)-Mn(2+)- H(3)PO(4)-luminol. The chemiluminescence intensity depends linearly on the logarithm of the concentration of human IgG over the range of 0.02-50ngml(-1) and detection limit (3sigma) is 0.005ngml(-1) (i.e., approximately 3x10(-14)M, 3amol in 100-mul sample). This assay has been successfully applied to the determination of human IgG in human serum samples and showed great potential for numerous applications in immunoassay.     

Biotechnological synthesis of Pd/Ag and Pd/Au nanoparticles for enhanced Suzuki–Miyaura cross-coupling activity

Bimetallic nanoparticle catalysts have attracted considerable attention due to their unique chemical and physical properties. The ability of metal-reducing bacteria to produce highly catalytically active monometallic nanoparticles is well known; however, the properties and catalytic activity of bimetallic nanoparticles synthesized with these organisms is not well understood. Here, we report the one-pot biosynthesis of Pd/Ag (bio-Pd/Ag) and Pd/Au (bio-Pd/Au) nanoparticles using the metal-reducing bacterium, Shewanella oneidensis, under mild conditions. Energy dispersive X-ray analyses performed using scanning transmission electron microscopy (STEM) revealed the presence of both metals (Pd/Ag or Pd/Au) in the biosynthesized nanoparticles. X-ray absorption near-edge spectroscopy (XANES) suggested a significant contribution from Pd(0) and Pd(II) in both bio-Pd/Ag and bio-Pd/Au, with Ag and Au existing predominately as their metallic forms. Extended X-ray absorption fine-structure spectroscopy (EXAFS) supported the presence of multiple Pd species in bio-Pd/Ag and bio-Pd/Au, as inferred from Pd–Pd, Pd–O and Pd–S shells. Both bio-Pd/Ag and bio-Pd/Au demonstrated greatly enhanced catalytic activity towards Suzuki–Miyaura cross-coupling compared to a monometallic Pd catalyst, with bio-Pd/Ag significantly outperforming the others. The catalysts were very versatile, tolerating a wide range of substituents. This work demonstrates a green synthesis method for novel bimetallic nanoparticles that display significantly enhanced catalytic activity compared to their monometallic counterparts.     

Radiative decay engineering 4. Experimental studies of surface plasmon-coupled directional emission

Fluorescence is typically isotropic in space and collected with low efficiency. In this paper we describe surface plasmon-coupled emission (SPCE), which displays unique optical properties and can be collected with an efficiency near 50%. SPCE occurs for fluorophores within about 200 nm of a thin metallic film, in our case a 50-nm-thick silver film on a glass substrate. We show that fluorophore proximity to this film converts the normally isotropic emission into highly directional emission through the glass substrate at a well-defined angle from the normal axis. Depending on the thickness of the polyvinyl alcohol (PVA) film on the silver, the coupling efficiency of sulforhodamine 101 in PVA ranged from 30 to 49%. Directional SPCE was observed whether the fluorophore was excited directly or by the evanescent field due to the surface plasmon resonance. The emission is always polarized perpendicular to the plane of incidence, irrespective of the polarization of the incident light. The lifetimes are not substantially changed, indicating a mechanism somewhat different from that observed previously for the effects of silver particles on fluorophores. Remarkably, the directional emission shows intrinsic spectral resolution because the coupling angles depend on wavelength. The distances over which SPCE occurs, 10 to 200 nm, are useful because a large number of fluorophores can be localized within this volume. The emission of more distant fluorophores does not couple into the glass, allowing background suppression from biological samples. SPCE can be expected to become rapidly useful in a variety of analytical and medical sensing applications.     

Controlled electrophoretic deposition of multifunctional nanomodules for bioelectrochemical applications

The design of an electrochemical glucose sensing device formed by the electrodeposition of multifunctional Au nanoparticles is reported here as a novel concept for an enhanced generic sensing platform. Initially gold nanoparticles (Au) were alternatively coated with a layer of positively charged redox polymer (ORP) and a negatively charged glucose oxidase (GOX) layer alternatively using layer-by-layer methodology to form multifunctional Au/ORP/GOX/ORP particles. The modification and stability of the Au nanoparticles was monitored by using UV-vis spectroscopy and zeta-potential measurements. The modified Au nanoparticles were electrophoretically deposited onto an electrode to produce an electrochemical glucose sensing device. A considerable influence of electrophoretic deposition time and potential was found on the sensing platform response. Preliminary responses to glucose addition showed an enhanced performance by applying an electrophoretic deposition potential of +1.2V vs. Ag/AgCl for 30min. The observed response in the case of microelectrode geometry was in the range of mAcm(2). This work also shows that the presence of a second outer ORP layer on the functionalised Au nanoparticles improved the response.     

Enhanced detection sensitivity of pegylated CdSe/ZnS quantum dots-based prostate cancer biomarkers by surface plasmon-coupled emission

We demonstrate the fabrication and detection of quantum dots (QDs)-based prostate specific antigens (PSAs) cancer protein biochips by using enhanced surface plasmon-coupled emission measurements (SPCE). The PSAs are immobilized on a SiO(2)-protected thin gold substrate and pegylated QDs which conjugated with antibodies of PSA are used as fluorescent probes. Due to the excellent brightness of the QDs and the high directionality of emission, as well as the high light collection efficiency of SPCE, the limit of detection (LOD) is down to 10 fg/mL (equal to 0.3 fM) for the PSA chips by using QDs-based cancer protein. We expect that this QDs-based SPCE measurement system with the low LOD supplies a great potential for detecting various cancer biomarkers that are present in only low concentrations within the human body.     

Adverse immune reactions to gold. I. Chronic treatment with an Au(I) drug sensitizes mouse T cells not to Au(I), but to Au(III) and induces autoantibody formation

Upon weekly i.m. injections of disodium gold thiomalate (Na2AuTM) 100% of A.SW mice produced IgG autoantibodies to antinuclear Ag and nucleolar Ag, respectively; about 70% of C57BL/6 mice produced IgG antinuclear Ag, whereas DBA/2 mice were resistant. Moreover, C57BL/6 mice, but not DBA/2 mice, showed increased mesangial deposits of IgG. These alterations were due not to disodium thiomalate, but to the gold ion of Na2AuTM. An assumed T cell reactivity of susceptible mouse strains to Na2AuTM was tested by means of the direct popliteal lymph node (PLN) assay. However, no distinct PLN reaction to Na2AuTM was detectable. Likewise, AuCl did not induce a PLN reaction. Both Na2AuTM and AuCl contain gold in the Au(I) state. The poor PLN responses to Au(I) contrasted with the strong PLN responses to Au(III) compounds. PLN reactions to Au(III) were dose dependent, T cell dependent, and specific. When Au(III) was reduced to Au(I) by addition of Na2TM or methionine before testing in the PLN assay its sensitizing capacity was significantly decreased. Thus, the oxidation state of gold, i.e., Au(III) vs Au(I), plays a major role for its sensitizing capacity. Therefore, we propose that the Au(I) of Na2AuTM is oxidized to Au(III) before T cells are sensitized and adverse immunologic reactions develop. Results obtained with the adoptive transfer PLN assay indicated that, indeed, repeated i.m. injections of Na2AuTM sensitized A.SW and C57BL/6 splenic T cells to Au(III).