Radiative decay engineering 3. Surface plasmon-coupled directional emission

A new method of fluorescence detection that promises to increase sensitivity by 20- to 1000-fold is described. This method will also decrease the contribution of sample autofluorescence to the detected signal. The method depends on the coupling of excited fluorophores with the surface plasmon resonance present in thin metal films, typically silver and gold. The phenomenon of surface plasmon-coupled emission (SPCE) occurs for fluorophores 20-250 nm from the metal surface, allowing detection of fluorophores over substantial distances beyond the metal-sample interface. SPCE depends on interactions of the excited fluorophore with the metal surface. This interaction is independent of the mode of excitation; that is, it does not require evanescent wave or surface-plasmon excitation. In a sense, SPCE is the inverse process of the surface plasmon resonance absorption of thin metal films. Importantly, SPCE occurs over a narrow angular distribution, converting normally isotropic emission into easily collected directional emission. Up to 50% of the emission from unoriented samples can be collected, much larger than typical fluorescence collection efficiencies near 1% or less. SPCE is due only to fluorophores near the metal surface and may be regarded as emission from the induced surface plasmons. Autofluorescence from more distal parts of the sample is decreased due to decreased coupling. SPCE is highly polarized and autofluorescence can be further decreased by collecting only the polarized component or only the light propagating with the appropriate angle. Examples showing how simple optical configurations can be used in diagnostics, sensing, or biotechnology applications are presented. Surface plasmon-coupled emission is likely to find widespread applications throughout the biosciences.     

Multi-wavelength immunoassays using surface plasmon-coupled emission

We describe a new method for multi-wavelength immunoassays using surface plasmon-coupled emission (SPCE). This phenomenon is coupling of excited fluorophores with a nearby thin metal film, in our case silver, resulting in strongly directional emission into the underlying glass substrate. The angle at which the radiation propagate through the prism depends on the surface plasmon angle for the relevant wavelength. These angles depend on emission wavelength, allowing measurement of multiple analytes using multiple emission wavelengths. We demonstrated this possibility using antibodies labeled with either Rhodamine Red-X or AlexaFluor 647. These antibodies were directed against an antigen protein bound to the silver surface. The emission from each labeled antibody occurred at a different angle on the glass prism, allowing independent measurement of surface binding of each antibody. This method of SPCE immunoassays can be readily extended to 4 or more wavelengths.     

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.     

Radiative decay engineering 7: Tamm state-coupled emission using a hybrid plasmonic–photonic structure

There is a continuing need to increase the brightness and photostability of fluorophores for use in biotechnology, medical diagnostics, and cell imaging. One approach developed during the past decade is to use metallic surfaces and nanostructures. It is now known that excited state fluorophores display interactions with surface plasmons, which can increase the radiative decay rates, modify the spatial distribution of emission, and result in directional emission. One important example is surface plasmon-coupled emission (SPCE). In this phenomenon, the fluorophores at close distances from a thin metal film, typically silver, display emission over a small range of angles into the substrate. A disadvantage of SPCE is that the emission occurs at large angles relative to the surface normal and at angles that are larger than the critical angle for the glass substrate. The large angles make it difficult to collect all of the coupled emission and have prevented the use of SPCE with high-throughput and/or array applications. In the current article, we describe a simple multilayer metal–dielectric structure that allows excitation with light that is perpendicular (normal) to the plane and provides emission within a narrow angular distribution that is normal to the plane. This structure consists of a thin silver film on top of a multilayer dielectric Bragg grating, with no nanoscale features except for the metal or dielectric layer thicknesses. Our structure is designed to support optical Tamm states, which are trapped electromagnetic modes between the metal film and the underlying Bragg grating. We used simulations with the transfer matrix method to understand the optical properties of Tamm states and localization of the modes or electric fields in the structure. Tamm states can exist with zero in-plane wavevector components and can be created without the use of a coupling prism. We show that fluorophores on top of the metal film can interact with the Tamm state under the metal film and display Tamm state-coupled emission (TSCE). In contrast to SPCE, the Tamm states can display either S or P polarization. The TSCE angle is highly sensitive to wavelength, which suggests the use of Tamm structures to provide both directional emission and wavelength dispersion. Metallic structures can modify fluorophore decay rates but also have high losses. Photonic crystals have low losses but may lack the enhanced light-induced fields near metals. The combination of plasmonic and photonic structures offers the opportunity for radiative decay engineering to design new formats for clinical testing and other fluorescence-based applications.     

Directional surface plasmon-coupled emission: A new method for high sensitivity detection

Fluorescence emission is nearly isotropic in space. With typical optical components the collection efficiency is 1% or less. In this preliminary report, we describe a novel approach to transforming the normally isotropic emission into directional emission with a collection efficiency near 50%. This can be accomplished for fluorophores located near a semi-transparent silver film on a glass substrate. The emission couples with the surface plasmon resonance on the silver surface and enters the transparent substrate at a sharply defined angle, the surface plasmon angle for the emission wavelength. We estimate that 40-70% of the total emission enters the substrate at the plasmon angle and can thus be directed towards a detector. Background emission from fluorophores distant from the silver does not couple with the plasmon and is not detected. Different emission wavelengths couple at different angles allowing spectral discrimination without additional optics. Surface plasmon-coupled emission represents a new technology which can be used for high detection efficiency with microfluidic and/or surface-bound assay formats.     

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.     

Directional surface plasmon-coupled emission from a 3 nm green fluorescent protein monolayer

High-sensitivity detection schemes are of great interest for a number of applications. Unfortunately, such schemes are usually high-cost. We demonstrate a low-cost approach to a high-sensitivity detection scheme based on surface plasmon-coupled emission (SPCE). The SPCE of a monomolecular layer of green fluorescent protein (GFP) is reported here. The protein was electrostatically attached to a thin, SiO(2)-protected silver film deposited on a quartz substrate. The visible, directional emission of GFP was observed at a sharp, well-defined angle of 47.5 degrees from the normal to the coupling prism, and the spectrum corresponded to that of GFP. The SPCE resulting from the reverse Kretschmann configuration showed a 12-fold enhancement over the free space fluorescence. The directional emission was 97% p-polarized. The directionality and high polarization can be coupled with the intrinsic spectral resolution of SPCE to be used in the design miniaturized spectrofluorometers. The observation of SPCE in the visible region of the spectrum from a monolayer of protein opens up new possibilities in protein-based sensing.     

Signal enhancement of surface plasmon-coupled emission (SPCE) with the evanescent field of surface plasmons on a bimetallic paraboloid biochip

We present a novel approach to the enhancement of surface plasmon-coupled emission (SPCE) using surface plasmon excitation in a bimetal (Ag/Au) layer and we validate the enhancement by presenting the results of a model human IgG immunoassay. Theoretical calculations using Fresnel's equations have been carried out to determine the optimum bimetallic composition and the resulting electric field enhancement. Signal enhancement of SPCE was confirmed using a range of bimetallic layers which were deposited on the surface of a high collection efficiency polymer array biochip and subsequently immobilized with Alexa Fluor 647 labeled anti-human IgG. The bimetallic film of Ag/Au (36/10nm) was determined as an optimum substrate for maximum SPCE signal which was a compromise between the long-term stability of the metal layer and the optimized evanescent field enhancement. An enhanced dose-dependent response was also demonstrated which was ～3 times greater than that detected with a pure gold layer. A human IgG immunoassay showed a dose-dependent response yielding a limit of detection of 1pg/ml by the 3σ rule. The improved performance of the bimetal layer compared to that of an assay carried out on a pure gold layer is attributed to the enhanced evanescent field intensity of surface plasmons in the bimetal combination which excites more fluorescence hence producing an enhanced SPCE signal. This result demonstrates the potential of the SPCE-based array biochips as a sensitive and high-throughput analysis platform for biomolecular interactions.     

Plasmon Hybridization in Silver Nanoislands as Semishells Arrays Coupled to a Thin Metallic Film

We obtained experimentally strong plasmon interactions between localized surface plasmon with delocalized surface plasmon polaritons in a new nanosystem of silver semishells island film arrays arranged as a closed-packing structure coupled to an adjacent thin silver film. We show that plasmon interactions for such a nanosystem exhibits two pronounced resonances and interpret the coupling in terms of Fano resonances. The higher energy resonance is identified as a symmetric hybridization mode between localized plasmon resonances in the island semishell array and surface plasmon polaritons in the metal film and while the lower energy resonance is identified as a corresponding anti-symmetric hybridization mode. Increasing the size of the particle arrays enhances and red shifts the resonances. We show that adding a dielectric spacer between the semishell island array and the metal film results in a red shifting of the resonances and introduce an additional high energy spectral peak. The effect of the spacer layer is interpreted as a reduced hybridization and the generation of additional localized surface plasmon resonances.     

Solution-Deposited Thin Silver Films on Plastic Surfaces for Low-Cost Applications in Plasmon-Coupled Emission Sensors

We report the deposition of highly uniform thin silver films on plastic materials using a wet-chemistry method, suitable for surface plasmon-coupled emission (SPCE). This approach is reproducible for diverse low-cost applications and versatile to generate silver surfaces on various plastics substrates. An oxygen plasma pretreatment of the plastic provides for rapid silvering, leading to a 47-nm-thick continuous film for SPCE applications. The surface smoothness and thickness of the films have been estimated using atomic force microscope. The higher refractive index of polycarbonate, resulted in an SPCE angle of θ _F = 47⁰ for Rhodamine B, compared to glass (θ _F = 50⁰). The current study presents details on film deposition conditions, appropriate choice of index matching fluids, substrates, and light sources that play a vital role to augment SPCE emission intensity.     

Surface plasmon resonance imaging on a microchip for detection of DNA-modified gold nanoparticles deposited onto the surface in a non-cross-linking configuration

We report theoretical predictions and experimental observations of the reduced detection volume with the use of surface-plasmon-coupled emission (SPCE). The effective fluorescence volume (detection volume) in SPCE experiments depends on two near-field factors: the depth of evanescent wave excitation and a distance-dependent coupling of excited fluorophores to the surface plasmons. With direct excitation of the sample (reverse Kretschmann excitation) the detection volume is restricted only by the distance-dependent coupling of the excitation to the surface plasmons. However, with the excitation through the glass prism at surface plasmon resonance angle (Kretschmann configuration), the detection volume is a product of evanescent wave penetration depth and distance-dependent coupling. In addition, the detection volume is further reduced by a metal quenching of excited fluorophores at a close proximity (below 10nm). The height of the detected volume size is 40-70nm, depending on the orientation of the excited dipoles. We show that, by using the Kretschmann configuration in a microscope with a high-numerical-aperture objective (1.45) together with confocal detection, the detection volume can be reduced to 1-2attoL. The strong dependence of the coupling to the surface plasmons on the orientation of excited dipoles can be used to study the small conformational changes of macromolecules.     

DNA hybridization assays using metal-enhanced fluorescence

We describe a new approach to DNA hybridization assays using metal-enhanced fluorescence. Thiolated oligonucleotides were bound to silver particles on a glass substrate. Addition of a complementary fluorescein-labeled oligonucleotide resulted in a dramatic time-dependent 12-fold increase in fluorescence intensity during hybridization. Proximity to silver particles resulted in a decreased fluorescence lifetime. This effect is thought to be the result of enhanced fluorescence from fluorescein near metallic silver particles. Hybridization could thus be measured from the decay kinetics of the emission, which can be measured independently from the emission intensity. These results suggest the use of silver particles as a general approach to measure DNA hybridization as a method to increase the sensitivity of DNA detection.     

Simultaneous Excitation of Surface Plasmon Polaritons on Both Interfaces in an Asymmetric Dielectric-Metal Corrugated Structure

The simultaneous excitation of plasmon polaritons on both surfaces of metal film was studied for asymmetric dielectric-metal-dielectric corrugated structures. Due to the small resonant absorption of the incident light on the transmission side of the structure, we investigated the enhancement of the surface plasmon polaritons on the mentioned side by controlling the structure parameters. When the illuminate light changes from normal incidence to non-normal incidence, the resonant absorption peak splits into a doublet. The simultaneous excitation of surface plasmon polaritons on both surfaces of the metal film can be achieved by controlling the incident angle. Since the wave vector matching condition is not satisfied, there is no coupling between the plasmon polaritons modes on the two surfaces of the corrugated metallic film. The excitation and control of the non-coupled surface plasmon polartions simultaneously propagating on the different interfaces of one metallic film have potential applications for designing novel compact and tunable nano-photonic devices at visible frequency.     

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.     

Tunable Extraordinary Optical Transmission in a Metal Film Perforated with Two-Level Subwavelength Cylindrical Holes

We propose a novel plasmonic metal structure composed of a silver film perforated with a two-dimensional square array of two-level cylindrical holes on a silica substrate. The transmission properties of this structure are theoretically calculated by the finite-difference time-domain (FDTD) method. Double-enhanced transmission peaks are achieved in the visible and infrared regions, which mainly originate from the excitation of localized surface plasmon resonances (LSPRs), the hybridization of plasmon modes, and the optical cavity mode formed in the holes. The enhanced transmission behaviors can be effectively tailored by changing the geometrical parameters and dielectric materials filled in the holes. These findings indicate that our proposed structure has potential applications in highly integrated optoelectronic devices.     

Excitation and Optimization Modeling of Surface Plasmon Polaritons in a Concentric Circular Metallic Grating Film

Interaction behavior between surface plasmon polaritons (SPPs) and Hankel-distributed diffracted waves (DWs) on a silver concentric circular grating film is studied using a rigorous coupled-wave technique for circular structure. It is shown that the numerical technique reveals the excitation characteristics of SPPs in the circular metal grating as well as provides an accurate calculation of SPP intensities for further optimization designs. Results show that the SPPs can be excited by various DWs through the control of wavelength and angle of the incident light. The most efficient excitation of SPPs from this circular metal grating structure can be obtained from the +1^st-order DW under a normal incidence with wavelength close to the grating period, and the optimal thickness and duty cycle of the grating are found to be 370 and 0.5 nm, respectively. It is shown that the optimized intensity of SPPs excited from circular metal grating can be higher than that from strip metal grating by over one order of magnitude.     

Surface Plasmon Polariton Modes at Interface of a Metal and an Ambichiral Sculptured Thin Film

The excitation of surface plasmon polariton (SPP) at interface of a metal and an ambichiral sculptured thin film was theoretically investigated in the Kretschmann configuration using the transfer matrix method. The dependence of SPP modes for a P polarization plane wave on the incident angle of light and the angle of rise of nanocolumns of ambichiral dielectric medium was reported. We found that multiple SPP modes are excited at the interface of metal and ambichiral dielectric medium. The results of phase speed as a function of pitch showed only that a SPP mode can be excited at all pitches.     

Long Range Surface Plasmons for Observation of Biomolecular Binding Events at Metallic Surfaces

A long range surface plasmon (LRSP) is an electromagnetic wave propagating along a thin metal film with an order of magnitude lower damping than conventional surface plasmon (SP) waves. Thus, the excitation of LRSP is associated with a narrower resonance and it provides larger enhancement of intensity of the electromagnetic field. In surface plasmon resonance (SPR) biosensors, these features allow a more precise observation of the binding of biomolecules in the proximity to the metal surface by using the (label-free) measurement of refractive index (RI) variations and by SP-enhanced fluorescence spectroscopy. In this contribution, we investigate LRSPs excited on a layer structure consisting of a fluoropolymer buffer layer, a thin gold film, and an aqueous sample. By implementing such structure in an SPR sensor, we achieved a 2.4- and 4.4-fold improvement of the resolution in the label-free and fluorescence-based detection, respectively, of the binding of biomolecules in the close proximity to the surface. Moreover, we demonstrate that the sensor resolution can be improved by a factor of 14 and 12 for the label-free and fluorescence-based detection, respectively, if the biomolecular binding events occur within the whole evanescent field of LRSP.     

A Surface Design for Enhancement of Light Trapping Efficiencies in Thin Film Silicon Solar Cells

We report on a surface design of thin film silicon solar cells based on silver nanoparticle arrays and blazed grating arrays. The light transmittance is increased at the front surface of the cells, utilizing the surface plasmon resonance effect induced by silver nanoparticle arrays. As a reflection layer structure, blazed gratings are placed at the rear surface to increase the light reflectance at bottom of the thin film cells. With the combination of the silver nanoparticle arrays and the blazed gratings, the light trapping efficiency of the thin film solar cell is characterized by its light absorptance, which is determined from the transmittance at front surface and the reflectance at bottom, via the finite-difference time-domain (FDTD) numerical simulation method. The results reveal that the light trapping efficiency is enhanced as the structural parameters are optimized. This work also shows that the surface plasmon resonance effect induced by the silver nanoparticles and the grating characteristics of the blazed gratings play crucial roles in the design of the thin film silicon solar cells.     

Optimization of Nanoparticle Size for Plasmonic Enhancement of Fluorescence

This paper reports on the enhancement of fluorescence that can result from the proximity of fluorophores to metallic nanoparticles (NPs). This plasmonic enhancement, which is a result of the localized surface plasmon resonance at the metal surface, can be exploited to improve the signal obtained from optical biochips and thereby lower the limits of detection. There are two distinct enhancement effects: an increase in the excitation of the fluorophore and an increase in its quantum efficiency. This study focuses on the first of these effects where the maximum enhancement occurs when the NP plasmon resonance wavelength coincides with the fluorophore absorption band. In this case, the excitation enhancement is proportional to the square of the amplitude of the electric field. The scale of the enhancement depends on many parameters, such as NP size and shape, metal type, and NP–fluorophore separation. A model system consisting of spherical gold/silver alloy NPs, surrounded by a silica spacer shell, to which is attached a fluorescent ruthenium dye, was chosen and the dependence of the fluorescence enhancement on NP diameter was investigated. Theoretical calculations, based on Mie theory, were carried out to predict the maximum possible enhancement factor for spherical NPs with a fixed composition and a range of diameters. Spherical NPs of the same composition were fabricated by chemical preparation techniques. The NPs were coated with a thin silica shell to overcome quenching effects and the dye was attached to the shell.