SERS detection of biomolecules using lithographed nanoparticles towards a reproducible SERS biosensor

In this paper we highlight the accurate spectral detection of bovine serum albumin and ribonuclease-A using a surface-enhanced Raman scattering (SERS) substrate based on gold nanocylinders obtained by electron-beam lithography (EBL). The nanocylinders have diameters from 100 to 180 nm with a gap of 200 nm. We demonstrate that optimizing the size and the shape of the lithographed gold nanocylinders, we can obtain SERS spectra of proteins at low concentration. This SERS study enabled us to estimate high enhancement factors (10(5) for BSA and 10(7) for RNase-A) of important bands in the protein Raman spectrum measured for 1 mM concentration. We demonstrate that, to reach the highest enhancement, it is necessary to optimize the SERS signal and that the main parameter of optimization is the LSPR position. The LSPR have to be suitably located between the laser excitation wavelength, which is 632.8 nm, and the position of the considered Raman band. Our study underlines the efficiency of gold nanocylinder arrays in the spectral detection of proteins.     

In situ Raman scattering study on a controllable plasmon-driven surface catalysis reaction on Ag nanoparticle arrays

Control of the plasmon-driven chemical reaction for the transformation of 4-nitrobenzenethiol to p,p'-dimercaptoazobenzene by Ag nanoparticle arrays was studied. The Ag nanoparticle arrays were fabricated by means of nanosphere lithography. By changing the PS particle size, the localized surface plasmon resonance (LSPR) peaks of the Ag nanoparticle arrays can be tailored from 460 to 560 nm. The controlled reaction process was monitored by in situ surface-enhanced Raman scattering. The reaction can be dramatically influenced by varying the duration of laser exposure, Ag nanoparticle size, laser power and laser excitation wavelength. The maximum reaction speed was achieved when the LSPR wavelength of the Ag nanoparticle arrays matched the laser excitation wavelength. The experimental results reveal that the strong LSPR can effectively drive the transfer of the 'hot' electrons that decay from the plasmon to the reactants. The experimental results were confirmed by theoretical calculations.     

Facile fabrication of gold nanoparticle arrays for efficient surface-enhanced Raman scattering

An effective and facile method for the fabrication of a surface-enhanced Raman scattering (SERS)-active film with closely packed gold nanoparticle (AuNP) arrays is proposed by self-assembly of different sizes (16, 25, 40 and 70?nm) of AuNPs at a toluene/water interface with ethanol as the inducer. The as-prepared AuNP arrays exhibit efficient Raman scattering enhancement, and the enhancement factors estimated using p-aminothiophenol as a probe molecule range from 10(5) to 10(7). This is attributed to the coupling electromagnetic SERS enhancement mechanism with additional localization field within closely packed AuNPs, which have greater SERS activity and reproducibility than that on aggregates and on self-assembled monolayers of isolated AuNPs on glass.     

Silver-island films as substrates for enhanced Raman scattering: effect of deposition rate on intensity.

The relationship between surface-enhanced resonance Raman scattering (SERRS) intensity and the rate of deposition during silver-island film preparation was examined, using zinc tetraphenylporphine (ZnTPP) as the adsorbate. The effect of the deposition rate on the optical properties of the films at specific wavelengths was also analyzed. The data show that the film extinction (the term extinction is used rather than absorption because the spectra have not been corrected for reflection or scattering losses) increases exponentially at 514 and 458 nm as the deposition rate is decreased. SERRS intensities also increase exponentially at these two excitation wavelengths with a decrease in the deposition rate. The optical density is linearly related to the SERRS intensity, and maximal enhancement is observed for films with the greatest extinction at these excitation wavelengths. In contrast, neither the extinction at 406 nm nor the SERRS scattering intensities resulting from excitation at this wavelength differ substantially. The surface-enhanced Raman scattering (SERS) intensity and the electronic spectra of 4,4'-bipyridine (BP) adsorbed onto silver films as a function deposition rate were also examined. The behavior of the nonresonantly enhanced BP is comparable to that of the resonantly enhanced ZnTPP samples. The effects of the surface morphology, as determined from transmission electron micrographs of the films at various deposition rates, on the corresponding electronic spectra and SERS/SERRS spectra are described.     

Surface-enhanced Raman scattering dendritic substrates fabricated by deposition of gold and silver on silicon

This paper reports a study on the preparation of gold nanoparticles and silver dendrites on silicon substrates by immersion plating. Firstly, gold was deposited onto silicon wafer from HF aqueous solution containing HAuCl4. Then, the silicon wafer deposited gold was dipped into HF aqueous solution of AgNO3 to form silver coating gold film. Scanning electron microscopy reveals a uniform gold film consisted of gold nanoparticles and rough silver coating gold film containing uniform dendritic structures on silicon surface. By SERS (surface-enhanced Raman scattering) measurements, the fabricated gold and silver coating gold substrates activity toward SERS is assessed. The SERS spectra of crystal violet on the fabricated substrates reflect the different SERS activities on gold nanoparticles film and silver coating gold dendrites film. Compared with pure gold film on silicon, the film of silver coating gold dendrites film significantly increased the SERS intensity. As the fabrication process is very simple, cost-effective and reproducible, and the fabricated silver coating gold substrate is of excellent enhancement ability, spatial uniformity and good stability.     

Vibrational spectra and surface-enhanced vibrational spectra of 1-nitropyrene

The present report on the vibrational spectra of 1-nitropyrene (1NP) describes the infrared and Raman spectra; their interpretation is aided by local density functional theory (DFT) calculations at the B3LYP/6-311G(d, p) level of theory and by the surface-enhanced vibrational spectra (SEVS) with the final objective of trace organic analytical applications. The surface-enhanced Raman scattering (SERS) on silver island films and mixed silver/gold island films was investigated with several laser lines in the visible region. Surface-enhanced infrared absorption (SEIRA) was attempted on silver and gold island films. The interface of the organic 1-NP with smooth metal surfaces of silver and copper was also probed using reflection-absorption infrared (RAIRS) spectra that, in conjunction with the transmission spectra, allow one to extract the molecular orientation in vacuum evaporated thin solid films. Chemical adsorption of 1-NP on silver and further photochemical decomposition of the 1-NP-metal adsorbates was detected with all visible laser lines. Resonance Raman scattering (RRS) using UV-laser excitation at 325 nm was also recorded.     

The relationship between extraordinary optical transmission and surface-enhanced Raman scattering in subwavelength metallic nanohole arrays

Nanohole arrays in an Ag film were used as a substrate for surface-enhanced Raman scattering in the optical range. Extraordinary optical transmission and local field enhancement in Ag nanohole arrays were theoretically simulated using three-dimensional finite difference time domain method. The periodicity of the holes was adjusted to control the transmission intensity and electric field intensity. The calculation results show that the peak position of transmission red-shifts as the periodicity increases, while the peak intensity decreases linearly. The electric field is localized in a very small region at the edges of the holes, which means the surface-enhanced Raman scattering originates only from a small number of molecules located in the edge regions. The electric field intensity changes with the excitation wavelength in a similar trend to the transmission intensity. Both the electric field intensity and transmission intensity reach their maximum value at the frequency of surface plasmon resonance. The structure that gives resonant transmission provides the maximum surface-enhanced Raman scattering signal. Controllable and predictable surface-enhanced Raman scattering can be produced by using this novel nanostructure. The structure can be optimized to get the maximum surface-enhanced Raman scattering signal at a certain excitation wavelength through numerical simulations.     

Shape-dependent surface-enhanced Raman scattering in gold-Raman probe-silica sandwiched nanoparticles for biocompatible applications

To meet the requirement of Raman probes (labels) for biocompatible applications, a synthetic approach has been developed to sandwich the Raman-probe (malachite green isothiocyanate, MGITC) molecules between the gold core and the silica shell in gold-SiO? composite nanoparticles. The gold-MGITC-SiO? sandwiched structure not only prevents the Raman probe from leaking out but also improves the solubility of the nanoparticles in organic solvents and in aqueous solutions even with high ionic strength. To amplify the Raman signal, three types of core, gold nanospheres, nanorods and nanostars, have been chosen as the substrates of the Raman probe. The effect of the core shape on the surface-enhanced Raman scattering (SERS) has been investigated. The colloidal nanostars showed the highest SERS enhancement factor while the nanospheres possessed the lowest SERS activity under excitation with 532 and 785 nm lasers. Three-dimensional finite-difference time domain (FDTD) simulation showed significant differences in the local electromagnetic field distributions surrounding the nanospheres, nanorods, and nanostars, which were induced by the localized surface plasmon resonance (LSPR). The electromagnetic field was enhanced remarkably around the two ends of the nanorods and around the sharp tips of the nanostars. This local electromagnetic enhancement made the dominant contribution to the SERS enhancement. Both the experiments and the simulation revealed the order nanostars > nanorods > nanospheres in terms of the enhancement factor. Finally, the biological application of the nanostar-MGITC-SiO? nanoparticles has been demonstrated in the monitoring of DNA hybridization. In short, the gold–MGITC-SiO? sandwiched nanoparticles can be used as a Raman probe that features high sensitivity, good water solubility and stability, low-background fluorescence, and the absence of photobleaching for future biological applications.     

Highly controlled surface-enhanced Raman scattering chips using nanoengineered gold blocks

Well defined gold nanostructures of various sizes are fabricated on glass substrates using high-resolution electron-beam lithography/lift-off techniques and detailed surface-enhanced Raman scattering (SERS) properties of crystal violet molecules are studied in order to elucidate electromagnetic (EM) field enhancement effects on the fabricated structures. SERS measurements are performed with high reproducibility using in situ Raman microspectroscopy in aqueous solution. An analysis based on EM theory is performed using field-enhancement factors obtained from finite-difference time-domain (FDTD) simulations and the analysis reproduces experimental results very well. It is noteworthy, furthermore, that the proposed analytic method of EM effects on SERS allows the estimate of the ideal local temperature of gold nanostructures by canceling out the difference in EM field factors at Stokes and anti-Stokes Raman scattering wavelengths. Thus, these experimental results demonstrate that quantitative analysis based on EM theory can be obtained using highly controlled gold nanostructures for SERS measurements with high reproducibility, a result that is promising for the construction of a SERS analysis chip. Although no SERS chip reported so far has been usable for quantitative analysis, this study opens the door for construction of a quantitative SERS chip.     

Surface-enhanced Raman spectroscopy to probe reversibly photoswitchable azobenzene in controlled nanoscale environments

We apply in situ surface-enhanced Raman spectroscopy (SERS) to probe the reversible photoswitching of azobenzene-functionalized molecules inserted in self-assembled monolayers that serve as controlled nanoscale environments. Nanohole arrays are fabricated in Au thin films to enable SERS measurements associated with excitation of surface plasmons. A series of SERS spectra are recorded for azobenzene upon cycling exposure to UV (365 nm) and blue (450 nm) light. Experimental spectra match theoretical calculations. On the basis of both the simulations and the experimental data analysis, SERS provides quantitative information on the reversible photoswitching of azobenzene in controlled nanoscale environments.     

Surface-enhanced vibrational spectroscopy of adsorbates on microemulsion synthesized gold nanoparticles

Very small (<10 nm) monodisperse gold nanoparticles (AuNPs) coated with a monolayer of decanethiol were prepared and their surface-enhanced infrared absorption (SEIRA) spectra were measured in the transmission mode. The AuNPs were prepared by the borohydride reduction of HAuCl(4) inside reverse micelles that were made by adding water to a hexane solution of sodium bis(2-ethylhexyl)sulfosuccinate (AOT). The gold nanoparticles were then stabilized by the addition of decanethiol. Subsequent addition of p-nitrothiophenol both facilitated the removal of excess AOT and showed that the gold surface was completely covered by the decanethiol. SEIRA spectra of decanethiol on gold particles prepared in AOT microemulsions were about twelve times more intense than corresponding layers on gold produced by electroless deposition and gave a significantly less noisy spectrum compared to the corresponding surface-enhanced Raman spectrum. The surface-enhanced Raman scattering (SERS) spectra of the same samples showed that the most intense spectrum was obtained from gold nanoparticles with a mean diameter of 2.5 nm. This result is in contrast to previous statements that SERS spectra could only be obtained from particles larger than 10 nm.     

Preparation of gold nano-cones as surface-enhanced Raman scattering sensors for molecule detection

Surface-enhanced Raman spectroscopy (SERS) is a powerful novel analytical tool which integrates high levels of sensitivity for trace analysis of chemical and biomolecular species due to the massive enhancement of Raman signals by using nanometre-sized metal particles. However, SERS can be envisaged as an analytical tool only if substrates with strong, predictable and reproducible SERS enhancement can be produced. Here we have developed one simple Ar+ ions sputtering technology to prepare gold nano-cones array on silicon substrates as surface-enhanced Raman scattering (SERS)-active substrates. The tip of the gold cone-structures exhibited an extremely sharp curvature with an apex diameter of 20 nm and the interior apex angle of the nanocones was around 20 degrees. These samples were evaluated as potential SERS substrates using Rhodamine 6G molecules as molecule probe and exhibited SERS enhancement factor of greater than 10, originated from the localized electron field enhancement around the apex of cones and the surface plasmon coupling of periodic structures.     

Plasmonic nanogap-enhanced Raman scattering using a resonant nanodome array

The optical properties and surface-enhanced Raman scattering (SERS) of plasmonic nanodome array (PNA) substrates in air and aqueous solution are investigated. PNA substrates are inexpensively and uniformly fabricated with a hot spot density of 6.25 × 10(6) mm(-2) using a large-area nanoreplica moulding technique on a flexible plastic substrate. Both experimental measurement and numerical simulation results show that PNAs exhibit a radiative localized surface plasmon resonance (LSPR) due to dipolar coupling between neighboring nanodomes and a non-radiative surface plasmon resonance (SPR) resulting from the periodic array structure. The high spatial localization of electromagnetic field within the ～10 nm nanogap together with the spectral alignment between the LSPR and excited and scattered light results in a reliable and reproducible spatially averaged SERS enhancement factor (EF) of 8.51 × 10(7) for Au-coated PNAs. The SERS enhancement is sufficient for a wide variety of biological and chemical sensing applications, including detection of common metabolites at physiologically relevant concentrations.     

Inverted size-dependence of surface-enhanced Raman scattering on gold nanohole and nanodisk arrays

Surface-enhanced Raman scattering (SERS) on gold nanohole and nanodisk arrays with precisely controlled size and spacing fabricated via electron beam lithography was investigated. These nanostructures exhibit strong SERS signals at 785 nm excitation but not at 514 nm. When the edge-to-edge distance is maintained, enhancement increases for nanoholes but decreases for nanodisks as diameter is increased. It is shown that the observed enhancement results from the local surface plasmon resonance wavelength shifts to the near-infrared regime as nanohole diameter increases. The large tolerance on dimensions and the empty space confined by nanoholes suggest promise for their use as a functional component in sensing, spectroscopy, and photonic devices.     

Realization of thermally durable close-packed 2D gold nanoparticle arrays using self-assembly and plasma etching

Realization of thermally and chemically durable, ordered gold nanostructures using bottom-up self-assembly techniques are essential for applications in a wide range of areas including catalysis, energy generation, and sensing. Herein, we describe a modular process for realizing uniform arrays of gold nanoparticles, with interparticle spacings of 2?nm and above, by using RF plasma etching to remove ligands from self-assembled arrays of ligand-coated gold nanoparticles. Both nanoscale imaging and macroscale spectroscopic characterization techniques were used to determine the optimal conditions for plasma etching, namely RF power, operating pressure, duration of treatment, and type of gas. We then studied the effect of nanoparticle size, interparticle spacing, and type of substrate on the thermal durability of plasma-treated and untreated nanoparticle arrays. Plasma-treated arrays showed enhanced chemical and thermal durability, on account of the removal of ligands. To illustrate the application potential of the developed process, robust SERS (surface-enhanced Raman scattering) substrates were formed using plasma-treated arrays of silver-coated gold nanoparticles that had a silicon wafer or photopaper as the underlying support. The measured value of the average SERS enhancement factor (2?×?10(5)) was quantitatively reproducible on both silicon and paper substrates. The silicon substrates gave quantitatively reproducible results even after thermal annealing. The paper-based SERS substrate was also used to swab and detect probe molecules deposited on a solid surface.     

Correct Spectral Conversion between Surface‐Enhanced Raman and Plasmon Resonance Scattering from Nanoparticle Dimers for Single‐Molecule Detection

Simultaneous measurement of surface‐enhanced Raman scattering (SERS) and localized surface plasmon resonance (LSPR) in nanoparticle dimers presents outstanding opportunities in molecular identification and in the elucidation of physical properties, such as the size, distance, and deformation of target species. SERS–LSPR instrumentation exists and has been used under limited conditions, but the extraction of SERS and LSPR readouts from a single measurement is still a challenge. Herein, the extraction of LSPR spectra from SERS signals is reported and a tool for measuring the interparticle distance from Raman enhancement data by the standardization of the SERS signal is proposed. The SERS nanoruler mechanism incorporates two important aspects (the LSPR scattering peak shift and the Raman shift for measuring interparticle distance), and signifies their exact one‐to‐one correspondence after spectral correction. The developed methodology is applied to calculate the interparticle distance between nanoparticle dimers from SERS signals, to detect and quantify DNA at the single‐molecule level in a base‐pair‐specific manner. It is also shown that the SERS nanoruler concept can be used in structural analysis for the specific detection of the interaction of immunoglobulin G (IgG) with its target from bianalyte Raman signals with identical shaping at single‐molecule resolution. The SERS profile shaping approach not only offers a new detection mechanism for single molecules, but also has excellent potential for studying protein interactions and the intracellular detection of mRNA.     

Probing intrinsic and extrinsic components in single osteosarcoma cells by near-infrared surface-enhanced Raman scattering

We report on the capabilities of near-infrared surface-enhanced Raman scattering (SERS) using gold nanoparticles to obtain detailed chemical information with high spatial resolution from within single cancer cells, living or fixed. Colloidal gold particles, 60 nm in size, were introduced into live human osteosarcoma cells by endocytosis by adding them to the growth medium. Rapid SERS mapping of cells indicated that not only could rich vibrational spectra be obtained from intrinsic cellular constituents both in the cytoplasm and nucleus and but also the distribution of extrinsic molecules introduced into the cells, in this case, rhodamine 6G could be characterized, suggesting that the intracellular distribution of chemotherapeutic agents could potentially be measured by this technique. We show that the SERS signal intensity from the cellular components increases and more spectral detail is acquired from dried cells when compared with hydrated cells in buffer. The data also show spectral fluctuations, mainly in intensity but also in peak position, which are dependent upon the intensity of the excitation light and are probably due to diffusion of molecules on the surface of the gold nanoparticles. A detailed understanding of the origins of these effects is still not complete, but the ability to acquire very sensitive SERS inside living cancer cells indicates the potential of this technique as a useful tool in biomedicine.     

One- and two-photon excited optical ph probing for cells using surface-enhanced Raman and hyper-Raman nanosensors

We demonstrate spatially resolved probing and imaging of pH in live cells by mobile and biocompatible nanosensors using surface-enhanced Raman scattering (SERS) of 4-mercaptobenzoic acid (pMBA) on gold nanoaggregates. Moreover, we also show that this concept of pH nanosensors can be extended to two-photon excitation by using surface-enhanced hyper-Raman scattering (SEHRS). In addition to the advantages of two-photon excitation, the SEHRS sensor enables measurements over a wide pH range without the use of multiple probes.     

Spherical Nanoparticle Arrays with Tunable Nanogaps and Their Hydrophobicity Enhanced Rapid SERS Detection by Localized Concentration of Droplet Evaporation

Periodic hexagonal spherical nanoparticle arrays are fabricated by a sacrificial colloidal monolayer template route by chemical deposition and further physical deposition. The regular network‐structured arrays are first templated by colloidal monolayers and then they are changed to novel periodic spherical nanoparticle arrays by further sputtering deposition due to multiple direction deposition and shadow effect between adjacent nanoparticles. The nanogaps between two adjacent spherical nanoparticles can be well tuned by controlling deposition time. Such periodic nanoparticle arrays with gold coatings demonstrate a very stable and high sensitive surface‐enhanced Raman scattering spectroscopy (SERS) performance. The periodic nanoparticle arrays with 10 nm gaps display much stronger SERS enhancement due to electromagnetic coupling. The chemically modified nanoparticle arrays show good hydrophobicity, which shorten process of detecting probe molecules using them as SERS‐active substrates by localized concentration of droplet evaporation and a low detection limit of 10⁻¹² m R6G can be achieved without solution wasting in a short time. The hydrophobic substrate offers a simple, convenient, and economical method to examine SERS performance by rapid concentration of solution on it and it is highly helpful to improve its practical applications in portable Raman detecting devices to detect organic molecules.     

In situ Raman monitoring of competitive adsorption of Ag and Au nanoparticles onto a poly(4-vinyl pyridine) surface

The competitive adsorption of citrate-capped Ag and Au nanoparticles (~25 nm in diameter) onto a poly(4-vinyl pyridine) (P4VP) surface has been investigated by means of Raman scattering spectroscopy. The P4VP film prepared on a glass slide was too thin for its normal Raman spectrum to be observed, but the Raman peaks of P4VP could be detected upon the adsorption of Ag and/or Au nanoparticles onto the film, due to the surface-enhanced Raman scattering (SERS) effect associated with the localized surface plasmon of Ag and/or Au nanoparticles. Neither quartz crystal microbalance nor atomic force microscopy (AFM) nor scanning electron microscopy (SEM) methodologies can distinguish between Ag and Au nanoparticles during their adsorption onto P4VP, but it is possible through Raman scattering spectroscopy because Ag (though not Au) nanoaggregates are SERS active at 514.5 nm excitation, while both Ag and Au nanoaggregates are SERS active at 632.8 nm excitation. Coupled with the AFM data, we were thus able to infer that about 120 Ag nanoparticles per 1 μm(2) were adsorbed, along with 60 Au nanoparticles per 1 μm(2), onto the P4VP film over a period of 1.5 h from a 1 : 1 mixture of Ag and Au sols at 1.6 nM each.