Study on the Mechanism of Diarylethene Crystal Growth by In Situ Microscopy and the Crystal Growth Controlled by an Aluminum Plasmonic Chip.

The microcrystalline film of an open-ring isomer (1o) of diarylethene 1 was prepared on an Al plasmonic chip with a grating structure. Photoisomerization from 1o to the closed-ring isomer (1c) and growth of needle-shaped crystals in 1c were observed in situ under an upright-inverted microscope. In the center part of the film, crystal growth of needle-shaped-crystal of 1c was observed upon UV irradiation from the top side, but not upon UV irradiation from the bottom side. However, crystallization occurred at the edge of the film upon UV irradiation from the bottom side. It was suggested that crystal growth of 1c required a high mobility of 1c near the film surface. Furthermore, the existence of 1o platform is also found to be required for alignment of 1c molecules by the results under the irradiation from the bottom and top sides. With the Al plasmonic chip, the conversion rate from 1o to 1c was larger inside the grating by the plasmonic enhanced field. Therefore, when the attenuated UV light was irradiated to the film edge with high mobility of 1c from the bottom side, the conversion rate was more than 60%, and the needle-shaped crystals of 1c were observed only inside the grating area. Crystal growth was controlled by the conversion rate of 1c promoted inside the grating. From the above, the larger conversion rate of 1c more than 60%, a high mobility of 1c near the film surface or edge, and the existence of the 1o platform for alignment of 1c molecules, are considered to be required for crystal growth in 1c.

Flexible Gold Nanocone Array Surfaces as a Tool for Regulating Neuronal Behavior.

Accelerated neurite outgrowth of rat cortical neurons on a flexible and inexpensive substrate functionalized with gold nanocone arrays is reported. The gold nanocone arrays are fabricated on Teflon films by a bottom-up approach based on colloidal lithography followed by deposition of a thin gold layer. The geometry of nanocone arrays including height and pitch is controlled by the overall etching time and template polystyrene beads size. Fluorescence microscopy studies reveal high viability and significant morphological changes of the neurons on the structured surfaces. The elongation degree of neurite is maximized on the nanocone arrays created with 1 µm polystyrene beads by a factor of two with respect to the control. Furthermore, the interface between the neurons and the nanocones is investigated by scanning electron microscopy and focused ion beam cross-sectioning. The detailed observation of the neuron/nanocone interfaces reveals the morphological similarity between the nanocone tips and the neuronal processes, the existence of interspace at the interface between the cell body and the nanocones, and neurite bridging among the neighboring structures, which may induce the acceleration of neurite outgrowth. The flexible gold nanocone arrays can be a good supporting substrate of neuron culture with noble electrical and optical properties.

Enhanced fluorescence microscopy with the Bull's eye-plasmonic chip.

A Bull's eye-plasmonic chip composed of concentric circles was applied to enhanced fluorescence microscopy. Among one dimensional (1-D), 2-D, and Bull's eye periodic structures, the Bull's eye-plasmonic chip provided the most enhanced fluorescence intensity under the epi-fluorescence microscope, because incident light through the objective lens with all azimuthal angles can be effectively applied to the surface plasmon resonance- field (excitation field) and the plasmon-enhanced emission was also effectively collected. In the fluorescence observation of a single nanoparticle, the enhanced fluorescence images for a microsphere with ϕ 2 µm and a nanosphere with ϕ 200 nm were observed. For the nanospheres with ϕ 40 and 20 nm, the fluorescence image, which was undetectable on a glass slide, was observed in a spatial resolution of roughly diffraction limit on the Bull's eye-plasmonic chip. Furthermore, the use of an appropriate pinhole at the aperture stop in the incident optical system improved the fluorescence enhancement. The applicability of a Bull's eye-plasmonic chip to fluorescence imaging was demonstrated.

Dual-Color Fluorescence Imaging of EpCAM and EGFR in Breast Cancer Cells with a Bull's Eye-Type Plasmonic Chip.

Surface plasmon field-enhanced fluorescence microscopic observation of a live breast cancer cell was performed with a plasmonic chip. Two cell lines, MDA-MB-231 and Michigan Cancer Foundation-7 (MCF-7), were selected as breast cancer cells, with two kinds of membrane protein, epithelial cell adhesion molecule (EpCAM) and epidermal growth factor receptor (EGFR), observed in both cells. The membrane proteins are surface markers used to differentiate and classify breast cancer cells. EGFR and EpCAM were detected with Alexa Fluor® 488-labeled anti-EGFR antibody (488-EGFR) and allophycocyanin (APC)-labeled anti-EpCAM antibody (APC-EpCAM), respectively. In MDA-MB231 cells, three-fold plus or minus one and seven-fold plus or minus two brighter fluorescence of 488-EGFR were observed on the 480-nm pitch and the 400-nm pitch compared with that on a glass slide. Results show the 400-nm pitch is useful. Dual-color fluorescence of 488-EGFR and APC-EpCAM in MDA-MB231 was clearly observed with seven-fold plus or minus two and nine-fold plus or minus three, respectively, on the 400-nm pitch pattern of a plasmonic chip. Therefore, the 400-nm pitch contributed to the dual-color fluorescence enhancement for these wavelengths. An optimal grating pitch of a plasmonic chip improved a fluorescence image of membrane proteins with the help of the surface plasmon-enhanced field.

Fabrication of broadband antireflective plasmonic gold nanocone arrays on flexible polymer films.

Flexible broadband antireflective and light-absorbing nanostructured gold thin films are fabricated by gold vapor deposition onto Teflon films modified with nanocone arrays. The nanostructures are created by the oxygen plasma etching of polystyrene bead monolayers on Teflon surfaces. The periodicity and height of the nanocone arrays are controlled by the bead diameter and the overall etching time. The gold nanocone arrays exhibit a reflectivity of less than 1% over a wide spectral range (450-900 nm) and a wide range of incident angles (0-70°); this unique optical response is attributed to a combination of diffractive scattering loss and localized plasmonic absorption. In addition to nanocones, periodic nanostructures of nanocups, nanopyramids, and nanocavities can be created by the plasma etching of colloidal bilayers. This fabrication method can be used to create flexible nanocone-structured gold thin films over large surface areas (cm(2)) and should be rapidly incorporated into new technological applications that require wide-angle and broadband antireflective coatings.

Direct detection of neuron-specific enolase using a spectrometer-free colorimetric plasmonic biosensor.

Sensitive detection of a tumor marker, neuron-specific enolase (NSE), was performed by a label-free direct immunoassay based on a colorimetric plasmonic biosensor. Reflective plasmonic colors of silver nanodome arrays provided a way for a sensitive refractive index sensor based on spectrometer-free colorimetric detection. The direct detection of NSE was demonstrated by a combination of a sensitive sensor substrate and image processing. The limit of detection (LOD) for NSE was determined to be 270 pM, which is lower than the clinical threshold value of NSE used for medical diagnostics of small-cell lung cancer. Since our substrate-based colorimetric plasmonic biosensor is compatible with smartphone detection, we believe that the presented biosensor will open up a way for biosensor technology for point-of-care testing as well as mobile health applications.

Surface plasmon-coupled emission on plasmonic Bragg gratings.

Surface plasmon-coupled emission (SPCE) from emitters in a close proximity to a plasmonic Bragg grating is investigated. In this study, the directional fluorescence emission mediated by Bragg-scattered surface plasmons and surface plasmons diffraction cross-coupled through a thin metallic film is observed by using the reverse Kretschmann configuration. We show that controlling of dispersion relation of these surface plasmon modes by tuning the refractive index at upper and lower interfaces of a dense sub-wavelength metallic grating enables selective reducing or increasing the intensity of the light emitted to certain directions. These observations may provide important leads for design of advanced plasmonic structures in applications areas of plasmon-enhanced fluorescence spectroscopy and nanoscale optical sources.

Collective plasmon modes excited on a silver nanoparticle 2D crystalline sheet.

This paper describes unique plasmonic characteristics of two dimensional (2D) crystalline sheets composed of homogeneous Ag nanoparticles (AgNPs) fabricated by the Langmuir-Schaefer method at an air-water interface. The localized surface plasmon resonance (LSPR) band of the Ag nanosheet was tuned by changing the interparticle distance of AgNPs via the length of the organic capping molecules. Red shift of the LSPR band of the AgNPs sheet followed an exponential law against the interparticle distance in a similar manner to the previous reports of metal nanodisc pairs. However, the shift was much larger and less dependent on the interparticle separation gap. This phenomenon is reasonably interpreted as the long-range interaction of LSPR in the 2D sheet ('delocalized' LSPR) confirmed by simulation using the finite difference time domain (FDTD) method. The FDTD simulation also revealed additional enhancement of local electric fields on the 2D sheet compared to those on the single or paired particles.

Nanoscale coupling of photons to vibrational excitation of Ag nanoparticle 2D array studied by scanning tunneling microscope light emission spectroscopy.

Scanning tunneling microscope light emission (STM-LE) spectroscopy has been utilized to elucidate the luminescence phenomena of Ag nanoparticles capped with myristate (myristate-capped AgNP) and 2-methyl-1-propanethiolate (C(4)S-capped AgNP) on the dodecanethiol-precovered Au substrate. The STM imaging revealed that myristate-capped AgNPs form an ordered hexagonal array whereas C(4)S-capped AgNPs show imperfect ordering, indicating that a shorter alkyl chain of C(4)S-capped AgNP is not sufficient to form rigid interdigitation. It should be noted that such a nanoparticle ordering affects the luminescence properties of the Ag nanoparticle. We found that the STM-LE is only detected from the Ag nanoparticles forming the two-dimensional superlattice. This indicates that the STM-LE of the Ag nanoparticle is radiated via the collective excitation of the local surface plasmon resonance (LSPR) spread over the Ag nanoparticles. Note that the STM-LE spectra of the Ag nanoparticles exhibit spike-like peaks superimposed on the broad light emission peak. Using Raman spectroscopy, we concluded that the spike-like structure appearing in the STM-LE spectra is associated with the vibrational excitation of the molecule embedded between Ag nanoparticles.

Plasmonic coloration of silver nanodome arrays for a smartphone-based plasmonic biosensor.

In this study, the utility of plasmonic coloration on silver nanodome arrays for sensitive and quantitative detection of biomolecules using a smartphone-based sensor is proposed. In particular, a quantitative analysis of DNA hybridization was achieved using the hue angle in the HSV color space obtained from a photograph of a sensing spot taken using a smartphone camera. Silver and gold nanodome arrays consisting of a polystyrene bead layer covered with a thin metal film can be created over a large area by a bottom-up fabrication process. The metal nanodome arrays exhibited unique colorations which can be tuned by the dome diameter ϕ, metal species, and refractive index of the surrounding medium. The measurement of the bulk refractive index sensitivity revealed that the Ag nanodome with ϕ = 500 nm can provide the highest sensitivity of up to 588 nm per refractive index unit. The detection of DNA hybridization was performed by using a bimetallic nanodome consisting of silver and thin gold overlayers and DNA modified gold nanoparticles (AuNPs) for enhancing the sensor signals. Upon the immobilization of AuNPs, the Ag nanodome (ϕ = 200 nm) exhibited a large shift in the resonance wavelength accompanied by a dramatic change in coloration. The analysis of detection sensitivity of DNA hybridization using a model system revealed that colorimetric detection based on hue can be used for the quantitative detection of biomolecules in the same manner as the spectroscopic method with a few pM level of detectable concentration.

Polydopamine Thin Films as Protein Linker Layer for Sensitive Detection of Interleukin-6 by Surface Plasmon Enhanced Fluorescence Spectroscopy.

Polydopamine (PDA) thin films are introduced to the surface modification of biosensor surfaces utilizing surface plasmon enhanced fluorescence spectroscopy (SPFS) as the linker layer of capture antibody on to the sensor surfaces. The capture antibody can be directly attached to the sensor surface without using any coupling agent by functionalizing the gold sensor surface with PDA thin films. The PDA coating is performed by a single-step preparation process by applying the dopamine solution on the sensor surface, which requires an extremely short incubation time (10 min). The real-time in situ measurement of the adsorption kinetics of the capture antibody onto the PDA-coated sensor surface is studied by surface plasmon resonance (SPR) spectroscopy. It reveals that the immobilization of capture antibody immediately occurs after introduction of a solution containing capture antibody, and the sensor surface is fully covered with the capture antibody. The sensitive detection of the cytokine marker interleukin-6 (IL-6) is performed by SPFS using a sandwich assay format with fluorescently labeled detection antibody. The sensor chips functionalized by PDA chemistry exhibited sensitive sensor responses with low nonspecific adsorption of the detection antibody onto the sensor surface. The detection limit of IL-6 with the developed SPFS biosensor is determined to be 2 pg/mL (100 fM), which is within the range of the diagnostic criteria. Our observation elucidates the remarkable utility of PDA coatings for chemical modification of the metallic sensor surfaces by a simple, brief, and inexpensive manner.

Flexible Teflon nanocone array surfaces with tunable superhydrophobicity for self-cleaning and aqueous droplet patterning.

Tunable hydrophobic/hydrophilic flexible Teflon nanocone array surfaces were fabricated over large areas (cm(2)) by a simple two-step method involving the oxygen plasma etching of a colloidal monolayer of polystyrene beads on a Teflon film. The wettability of the nanocone array surfaces was controlled by the nanocone array dimensions and various additional surface modifications. The resultant Teflon nanocone array surfaces were hydrophobic and adhesive (a "gecko" type of surface on which a water droplet has a high contact angle but stays in place) with a contact angle that correlated with the aspect ratio/sharpness of the nanocones. The surfaces switched to a superhydrophobic or "lotus" type of surface when hierarchical nanostructures were created on Teflon nanocones by modifying them with a gold nanoparticle (AuNPs) film. The nanocone array surfaces could be made superhydrophobic with a maximum contact angle of 160° by the further modification of the AuNPs with an octadecanethiol (C18SH) monolayer. Additionally, these nanocone array surfaces became hydrophilic when the nanocone surfaces were sequentially modified with AuNPs and hydrophilic polydopamine (PDA) layers. The nanocone array surfaces were tested for two potential applications: self-cleaning superhydrophobic surfaces and for the passive dispensing of aqueous droplets onto hybrid superhydrophobic/hydrophilic microarrays.

Plasmon-Enhanced Fluorescence Biosensors: a Review.

Surfaces of metallic films and metallic nanoparticles can strongly confine electromagnetic field through its coupling to propagating or localized surface plasmons. This interaction is associated with large enhancement of the field intensity and local optical density of states which provides means to increase excitation rate, raise quantum yield, and control far field angular distribution of fluorescence light emitted by organic dyes and quantum dots. Such emitters are commonly used as labels in assays for detection of chemical and biological species. Their interaction with surface plasmons allows amplifying fluorescence signal (brightness) that accompanies molecular binding events by several orders of magnitude. In conjunction with interfacial architectures for the specific capture of target analyte on a metallic surface, plasmon-enhanced fluorescence (PEF) that is also referred to as metal-enhanced fluorescence (MEF) represents an attractive method for shortening detection times and increasing sensitivity of various fluorescence-based analytical technologies. This review provides an introduction to fundamentals of PEF, illustrates current developments in design of metallic nanostructures for efficient fluorescence signal amplification that utilizes propagating and localized surface plasmons, and summarizes current implementations to biosensors for detection of trace amounts of biomarkers, toxins, and pathogens that are relevant to medical diagnostics and food control.

Active Control of SPR by Thermoresponsive Hydrogels for Biosensor Applications.

The use of thermoresponsive poly(N-isopropylacrylamide)-based hydrogel (pNIPAAm) for rapid tuning of surface plasmon resonance (SPR) is reported. This approach is implemented by using an SPR layer architecture with an embedded indium tin oxide microheater and pNIPAAm film on its top. It takes advantage of rapid thermally induced swelling and collapse of pNIPAAm that is accompanied by large refractive index changes and leads to high thermo-optical coefficient of dn/dT = 2 × 10-2 RIU/K. We show that this material is excellently suited for efficient control of refractive index-sensitive SPR and that it can serve simultaneously as a 3D binding matrix in biosensor applications (if modified with biomolecular recognition elements for a specific capture of target analyte). We demonstrate that this approach enables modulating of the output signal in surface plasmon-enhanced fluorescence spectroscopy biosensors and holds potential for simple time-multiplexing of sensing channels for parallelized readout of fluorescence assays.

Bloch surface wave-enhanced fluorescence biosensor.

A new approach to signal amplification in fluorescence-based assays for sensitive detection of molecular analytes is reported. It relies on a sensor chip carrying a one-dimensional photonic crystal (1DPC) composed of two piled up segments which are designed to increase simultaneously the excitation rate and the collection efficiency of fluorescence light. The top segment supports Bloch surface waves (BSWs) at the excitation wavelength and the bottom segment serves as a Bragg mirror for the emission wavelength of used fluorophore labels. The enhancement of the excitation rate on the sensor surface is achieved through the resonant coupling to BSWs that is associated with strong increase of the field intensity. The increasing of collection efficiency of fluorescence light emitted from the sensor surface is pursued by using the Bragg mirror that minimizes its leakage into a substrate and provides its beaming toward a detector. In order to exploit the whole evanescent field of BSW, extended three-dimensional hydrogel-based binding matrix that is functionalized with catcher molecules is attached to 1DPC for capturing of target analyte from a sample. Simulations supported by experiments are presented to illustrate the design and determined the performance characteristics of BSW-enhanced fluorescence spectroscopy. A model immunoassay experiment demonstrates that the reported approach enables increasing signal to noise ratio, resulting in about one order of magnitude improved limit of detection (LOD) with respect to regular total internal reflection fluorescence (TIRF) configuration.