A SERS-active microfluidic device with tunable surface plasmon resonances

A surface-enhanced Raman scattering (SERS)-active microfluidic device with tunable surface plasmon resonances is presented here. It is constructed by silver grating substrates prepared by two-beam laser interference of photoresists and subsequent metal evaporation coating, as well as PDMS microchannel derived from soft lithography. By varying the period of gratings from 200 to 550 nm, surface plasmon resonances (SPRs) from the metal gratings could be tuned in a certain range. When the SPRs match with the Raman excitation line, the highest enhancement factor of 2×10(7) is achieved in the SERS detection. The SERS-active microchannel with tunable SPRs exhibits both high enhancement factor and reproducibility of SERS signals, and thus holds great promise for applications of on-chip SERS detection.     

Surface-enhanced vibrational spectroscopy of B vitamins: what is the effect of SERS-active metals used?

Surface-enhanced Raman scattering (SERS) spectroscopy and surface-enhanced infrared absorption (SEIRA) spectroscopy are analytical tools suitable for the detection of small amounts of various analytes adsorbed on metal surfaces. During recent years, these two spectroscopic methods have become increasingly important in the investigation of adsorption of biomolecules and pharmaceuticals on nanostructured metal surfaces. In this work, the adsorption of B-group vitamins pyridoxine, nicotinic acid, folic acid and riboflavin at electrochemically prepared gold and silver substrates was investigated using Fourier transform SERS spectroscopy at an excitation wavelength of 1,064 nm. Gold and silver substrates were prepared by cathodic reduction on massive platinum targets. In the case of gold substrates, oxidation–reduction cycles were applied to increase the enhancement factor of the gold surface. The SERS spectra of riboflavin, nicotinic acid, folic acid and pyridoxine adsorbed on silver substrates differ significantly from SERS spectra of these B-group vitamins adsorbed on gold substrates. The analysis of near-infrared-excited SERS spectra reveals that each of B-group vitamin investigated interacts with the gold surface via a different mechanism of adsorption to that with the silver surface. In the case of riboflavin adsorbed on silver substrate, the interpretation of surface-enhanced infrared absorption (SEIRA) spectra was also helpful in investigation of the adsorption mechanism.     

Highly sensitive signal detection of duplex dye-labelled DNA oligonucleotides in a PDMS microfluidic chip: confocal surface-enhanced Raman spectroscopic study

Rapid and highly sensitive detection of duplex dye-labelled DNA sequences in a PDMS microfluidic channel was investigated using confocal surface enhanced Raman spectroscopy (SERS). This method does not need either an immobilization procedure or a PCR amplification procedure, which are essential for a DNA microarray chip. Furthermore, Raman peaks of each dye-labelled DNA can be easily resolved since they are much narrower than the corresponding broad fluorescence bands. To find the potential applicability of confocal SERS for sensitive bio-detection in a microfluidic channel, the mixture of two different dye-labelled (TAMRA and Cy3) sex determining Y genes, SRY and SPGY1, was adsorbed on silver colloids in the alligator teeth-shaped PDMS microfluidic channel and its SERS signals were measured under flowing conditions. Its major SERS peaks were observable down to the concentration of 10(-11) M. In the present study, we explore the feasibility of confocal SERS for the highly sensitive detection of duplex dye-labelled DNA oligonucleotides in a PDMS microfluidic chip.     

Fast and sensitive trace analysis of malachite green using a surface-enhanced Raman microfluidic sensor

A rapid and highly sensitive trace analysis technique for determining malachite green (MG) in a polydimethylsiloxane (PDMS) microfluidic sensor was investigated using surface-enhanced Raman spectroscopy (SERS). A zigzag-shaped PDMS microfluidic channel was fabricated for efficient mixing between MG analytes and aggregated silver colloids. Under the optimal condition of flow velocity, MG molecules were effectively adsorbed onto silver nanoparticles while flowing along the upper and lower zigzag-shaped PDMS channel. A quantitative analysis of MG was performed based on the measured peak height at 1615 cm⁻¹ in its SERS spectrum. The limit of detection, using the SERS microfluidic sensor, was found to be below the 1–2 ppb level and this low detection limit is comparable to the result of the LC-Mass detection method. In the present study, we introduce a new conceptual detection technology, using a SERS microfluidic sensor, for the highly sensitive trace analysis of MG in water.     

Surface enhanced Raman spectroscopy for microfluidic pillar arrayed separation chips

Numerous studies have addressed the challenges of implementing miniaturized microfluidic platforms for chemical and biological separation applications. However, the integration of real time detection schemes capable of providing valuable sample information under continuous, ultra low volume flow regimes has not fully been addressed. In this report we present a chip based chromatography system comprising of a pillar array separation column followed by a reagent channel for passive mixing of a silver colloidal solution into the eluent stream to enable surface enhanced Raman spectroscopy (SERS) detection. Our design is the first integrated chip based microfluidic device to combine pressure driven separation capability with real time SERS detection. With this approach we demonstrate the ability to collect distinctive SERS spectra with or without complete resolution of chromatographic bands. Computational fluidic dynamic (CFD) simulations are used to model the diffusive mixing behaviour and velocity profiles of the two confluent streams in the microfluidic channels. We evaluate the SERS spectral band intensity and chromatographic efficiency of model analytes with respect to kinetic factors as well as signal acquisition rates. Additionally, we discuss the use of a pluronic modified silver colloidal solution as a means of eliminating contamination generally caused by nanoparticle adhesion to channel surfaces.     

Silver nanoparticle-treated filter paper as a highly sensitive surface-enhanced Raman scattering (SERS) substrate for detection of tyrosine in aqueous solution

Highly sensitive SERS substrates based on deposition of silver nanoparticles on commercially available filter paper were prepared in this work, and used to overcome problems found in analyses of aqueous samples. To prepare silver nanoparticle- (AgNP) doped filter substrates, a silver mirror reaction was used. The procedures for substrate preparation were systematically optimized. Pretreatment of filter paper, reaction time, temperature, and concentration of reagents for silver mirror reactions were studied. The morphologies of the resulting substrates were characterized by field-emission scanning electron microscopy (FE-SEM) and correlated with the SERS signals by probing with p-nitrothiophenol (pNTP). Filter papers with different pretreatments were found to have different sizes and distributions of AgNPs. The best performance was found when filter paper was pre-treated with ammonia solution before growth of AgNPs. Based on the SEM images, the resulting AgNPs had roughly spherical shape with a high degree of uniformity. The silver-coated filter paper substrates provide much higher SERS signals compared to glass substrates and the reproducibility was improved significantly. Based on statistical analyses, the relative standard deviations for substrate-to-substrate and spot-to-spot were both were less than 8% and the enhancement factors for the substrates were, in general, higher than 107. The SERS substrates were used to selectively detect tyrosine in aqueous solution. Results indicate that filter-based SERS substrates are highly suited to detection of tyrosine. Compared to glass-based SERS substrates, 50 times more SERS signal was observed in detection of tyrosine. The linear range can be up to 100 μM with a detection limit of 625 nM (SN(-1)=3).     

Fabrication of bimetallic microfluidic surface-enhanced Raman scattering sensors on paper by screen printing

Au–Ag bimetallic microfluidic, dumbbell-shaped, surface enhanced Raman scattering (SERS) sensors were fabricated on cellulose paper by screen printing. These printed sensors rely on a sample droplet injection zone, and a SERS detection zone at either end of the dumbbell motif, fabricated by printing silver nanoparticles (Ag NPs) and gold nanoparticles (Au NPs) successively with microscale precision. The microfluidic channel was patterned using an insulating ink to connect these two zones and form a hydrophobic circuit. Owing to capillary action of paper in the millimeter-sized channels, the sensor could enable self-filtering of fluids to remove suspended particles within wastewater without pumping. This sensor also allows sensitive SERS detection, due to advantageous combination of the strong surface enhancement of Ag NPs and excellent chemical stability of Au NPs. The SERS performance of the sensors was investigated by employing the probe rhodamine 6G, a limit of detection (LOD) of 1.1 × 10⁻¹³ M and an enhancement factor of 8.6 × 10⁶ could be achieved. Moreover, the dumbbell-shaped bimetallic sensors exhibited good stability with SERS performance being maintained over 14 weeks in air, and high reproducibility with less than 15% variation in spot-to-spot SERS intensity. Using these dumbbell-shaped bimetallic sensors, substituted aromatic pollutants in wastewater samples could be quantitatively analyzed, which demonstrated their excellent capability for rapid trace pollutant detection in wastewater samples in the field without pre-separation.     

Surface-enhanced Raman scattering: realization of localized surface plasmon resonance using unique substrates and methods

Surface-enhanced Raman scattering (SERS) enhancement and the reproducibility of the SERS signal strongly reflect the quality and nature of the SERS substrates because of diverse localized surface plasmon resonance (LSPR) excitations excited at interstitials or sharp edges. LSPR excitations are the most important ingredients for achieving huge enhancements in the SERS process. In this report, we introduce several gold and silver nanoparticle-based SERS-active substrates developed solely by us and use these substrates to investigate the influence of LSPR excitations on SERS. SERS-active gold substrates were fabricated by immobilizing colloidal gold nanoparticles on glass slides without using any surfactants or electrolytes, whereas most of the SERS-active substrates that use colloidal gold/silver nanoparticles are not free of surfactant. Isolated aggregates, chain-like elongated aggregates and two-dimensional (2D) nanostructures were found to consist mostly of monolayers rather than agglomerations. With reference to correlated LSPR and SERS, combined experiments were carried out on a single platform at the same spatial position. The isolated aggregates mostly show a broadened and shifted SPR peak, whereas a weak blue-shifted peak is observed near 430 nm in addition to broadened peaks centered at 635 and 720 nm in the red spectral region in the chain-like elongated aggregates. In the case of 2D nanostructures, several SPR peaks are observed in diverse frequency regions. The characteristics of LSPR and SERS for the same gold nanoaggregates lead to a good correlation between SPR and SERS images. The elongated gold nanostructures show a higher enhancement of the Raman signal than the the isolated and 2D samples. In the case of SERS-active silver substrates for protein detection, a new approach has been adopted, in contrast to the conventional fabrication method. Colloidal silver nanoparticles are immobilized on the protein functionalized glass slides, and further SERS measurements are carried out based on LSPR excitations. A new strategy for the detection of biomolecules, particularly glutathione, under aqueous conditions is proposed. Finally, supramolecular J-aggregates of ionic dyes incorporated with silver colloidal aggregates are characterized by SERS measurements and correlated to finite-difference time-domain analysis with reference to LSPR excitations. Figure SPR and SERS images for isolated, elongated and two-dimensional gold nanostructures     

Quantitative surface enhanced Raman scattering detection based on the "sandwich" structure substrate

A sandwich structured substrate was designed for quantitative molecular detection using surface enhanced Raman scattering (SERS), in which the probe molecule was sandwiched between silver nanoparticles (SNPs) and silver nanoarrays. The SNPs was prepared using Lee-Meisel method, and the silver nanoarrays was fabricated on porous anodic aluminum oxide (AAO) using electrodepositing method. The SERS studies show that the sandwich structured substrate exhibits good stability and reproducibility, and the detection sensitivity of Rhodamine 6G (R6G) and Melamine can respectively reach up to 10(-19) M and 10(-9) M, which is improved greatly as compared to other SERS substrates. The improved SERS sensitivity is closely associated with the stronger electromagnetic field enhancement, which stems from localized surface plasmon (LSP) coupling between the two silver nanostructures. Furthermore, the SERS intensity increased almost linearly as the mother concentration increased, which indicates that such a sandwich structure may be used as a good SERS substrate for quantitative analysis.     

Microfluidic fabrication of SERS-active microspheres for molecular detection

In this paper, we demonstrated a microfluidic system for fabricating microspheres with hierarchical surface nanopatterns for molecular detection based on surface-enhanced Raman scattering (SERS). Briefly, a photocurable silica suspension was emulsified into monodisperse droplets using a microfluidic device composed of two coaxial glass capillaries. The silica particles in each droplet protruded through the interface and spontaneously formed a hexagonal array. After polymerization of the droplets, we selectively decorated the exposed areas of the silica particles with silver nanoparticles through electroless deposition. The resulting hierarchically-structured microspheres showed high sensitivity and fast binding kinetics in molecular detection based on SERS, owing to the dense array of hot spots on each microsphere and high mobility of the microspheres, respectively. Notably, the SERS signals from molecules adsorbed on the microspheres could be detected in both the dried and suspension states. In addition, we demonstrated that the SERS-active microspheres can be functionalized into structural colored or magnetoresponsive microspheres for advanced applications.     

Sub-wavelength Raman spectroscopy on isolated silver islands

Surface Enhanced Raman Spectroscopy (SERS) is a valuable analytical tool for the investigation of molecules adsorbed on roughened noble metal surfaces. The shape, size, and surrounding of the metal protrusions play an important role in the Raman scattering enhancement. By combining scanning near-field optical microscopy (SNOM) with Raman spectroscopy the spatial resolution suffices for investigating isolated silver islands on SERS active substrates. We demonstrate an optical resolution below 70 nm for recording spectra on specifically prepared and fully characterized SERS substrates. For a quantitative evaluation of the SERS signal the spatial distribution of Rhodamine 6G (R6G) deposited on the SERS substrate was determined by friction force measurements. By comparing the Raman intensities of the SERS substrates with those of unmetallized support plates absolute SERS enhancement factors at specific locations on top and in the vicinity of the silver islands were determined directly.     

Ultrastable substrates for surface-enhanced Raman spectroscopy: Al2O3 overlayers fabricated by atomic layer deposition yield improved anthrax biomarker detection

A new method to stabilize and functionalize surfaces for surface-enhanced Raman spectroscopy (SERS) is demonstrated. Atomic layer deposition (ALD) is used to deposit a sub-1-nm alumina layer on silver film-over-nanosphere (AgFON) substrates. The resulting overlayer maintains and stabilizes the SERS activity of the underlying silver while presenting the surface chemistry of the alumina overlayer, a commonly used polar adsorbent in chromatographic separations. The relative affinity of analytes for alumina-modified AgFON substrates can be determined by their polarity. On the basis of SERS measurements, dipicolinic acid displays the strongest binding to the ALD alumina-modified AgFON among a set of pyridine derivatives with varying polarity. This strong affinity for carboxylate groups makes the SERS substrate an ideal candidate for bacillus spores detection using the dipicolinate biomarker. The SERS signal from extracted dipicolinate was measured over the spore concentration range 10(-14)-10(-12) M to determine the saturation binding capacity of the alumina-modified AgFON surface. The adsorption constant was determined to be Kspore = 9.0 x 10(13) M(-1). A 10-s data collection time is capable of achieving a limit of detection of approximately 1.4 x 10(3) spores. The shelf life of prefabricated substrates is at least 9 months prior to use. In comparison to the bare AgFON substrates, the ALD-modified AgFON substrates demonstrate twice the sensitivity with 6 times shorter data acquisition time and 7 times longer temporal stability. ALD expands the palette of available chemical methods to functionalize SERS substrates, which will enable improved and diverse chemical control over the nature of analyte-surface binding for biomedical, homeland security, and environmental applications.     

Recent advances in surface-enhanced Raman scattering detection technology for microfluidic chips

Microfluidic chip devices and their application to sensitive chemical and biological analyses have attracted significant attention over the past decade. The miniaturization of reaction systems offers practical advantages over conventional benchtop systems. In this case, however, a highly sensitive on-chip detection method is important for the monitoring of chemical reactions as well as for the detection of analytes inside the channel because the detection volume in a micrometer-size channel is extremely small. Recently, a surface-enhanced Raman scattering (SERS) technique is being regarded as a potential candidate for the highly sensitive detection of analytes in a microfluidic chip. This review provides a general survey and an in-depth look at recent developments in SERS techniques for the biological/environmental analysis of minute analytes in a microfluidic chip.     

Surface-enhanced Raman scattering of a series of n-hydroxybenzoic acids (n = P, M and O) on the silver nano-particles

Surface-enhanced Raman scattering (SERS) spectra of a series of n-hydroxybenzoic acids (n-HBA, n = P, M and O) adsorbed on the silver nano-particles were studied, respectively, in the silver colloidal solution and on the dried silver-coated filter paper. On the same substrate, the different molecules' SERS spectra were different, while on the different substrates the same molecules' SERS spectra were also different. Significant changes were found in the SERS spectra of PHBA molecules adsorbed on the two substrates, and the changes found in MHBA's spectra on two substrates were next to PHBA's, while almost no changes were found in the spectra of OHBA molecules. Moreover, it was found, on the filter paper, that the SERS spectra of the same molecules would change with the coverage density of the silver nano-particles. The analyses showed that the origins of these changes were the different adsorption behavior of molecules adsorbed on the silver nano-particles. Because in these three molecules the relative positions of the carboxyls and hydroxyls on the benzenes are different, the adsorption behaviors of these three molecules adsorbed on the silver surfaces are also different. The experimental results suggest that the surface characteristic of the substrate and the surface configuration of the adsorbate could exert a great influence on the adsorption behavior of the adsorbates on the substrates.     

In situ laser-induced photochemical silver substrate synthesis and sequential SERS detection in a flow cell

A new, simple, and effective approach for multianalyte sequential surface-enhanced Raman scattering (SERS) detection in a flow cell is reported. The silver substrate was prepared in situ by laser-induced photochemical synthesis. By focusing the laser on the 320 μm inner diameter glass capillary at 0.5 ml/min continuous flow of 1 mM silver nitrate and 10 mM sodium citrate mixture, a SERS active silver spot on the inner wall of the glass capillary was prepared in a few seconds. The test analytes, dacarbazine, 4-(2-pyridylazo)resorcinol (PAR) complex with Cu(II), and amoxicillin, were sequentially injected into the flow cell. Each analyte was adsorbed to the silver surface, enabling the recording of high intensity SERS spectra even at 2 s integration times, followed by desorption from the silver surface and being washed away from the capillary. Before and after each analyte passed the detection window, citrate background spectra were recorded, and thus, no “memory effects” perturbed the SERS detection. A good reproducibility of the SERS spectra obtained under flow conditions was observed. The laser-induced photochemically synthesized silver substrate enables high Raman enhancement, is characterized by fast preparation with a high success rate, and represents a valuable alternative for silver colloids as SERS substrate in flow approaches.     

Electrochemical synthesis and functionality evaluation of silver nanostructured layers

The functionality of silver nanostructures prepared by means of electrochemical deposition of silver into the pores of anodic alumina oxide (AAO) template was examined in correlation to electrodeposition conditions. The optical activity as well as the chemical separation ability of prepared nanostructured films was studied. The surface enhanced Raman spectroscopy (SERS) performance was evaluated by the signals of rhodamine 6G, 4‐aminothiophenol and 2,7‐dichlorfluorescein. Nanostructured silver substrates showed moderate surface enhancement for Raman scattering from adsorbed molecules with the magnitude of about 26.9. Moreover, a novel separation/pre‐concentration function of the silver nanowire structures was indicated. The identification and position detection of the model compounds were realised with SERS. The separation of single chemical components from the two‐component mixture over the examined silver nanostructured films was sufficiently approved. The results obtained demonstrated the potential of the prepared substrate as a SERS detection and separation probe for further implementation to any instrumentation. Copyright © 2014 John Wiley & Sons, Ltd.     

The optimisation of facile substrates for surface enhanced Raman scattering through galvanic replacement of silver onto copper

A fast and cost-effective approach for the synthesis of substrates used in surface enhanced Raman scattering (SERS) has been developed using galvanic displacement. Deposition of silver onto commercially available Cu foil has resulted in the formation of multiple hierarchical structures, whose morphology show dependence on deposition time and temperature. Analysis of the surface structure by scanning electron microscopy revealed that the more complex silver structures correlated well with increased deposition time and temperature. Using Rhodamine 6G (R6G) as a model Raman probe it was also possible to relate the substrate morphology directly with subsequent SERS intensity from the R6G analyte as well as the reproducibility across a total of 15 replicate Raman maps (20 × 20 pixels) consisting of 400 spectra at a R6G concentration of 10(-4) M. The substrate with the highest reproducibility was then used to explore the limit of detection and this compared very favourably with colloidal-based SERS assessments of the same analyte.     

Synthesis and Characterization of Silver Nanoparticle Modified 3-Aminophenol Resin Microspheres with Application for Determination of Carcinoembryonic Antigens by Surface-Enhanced Raman Scattering

Uniform phenolic resin microspheres were prepared by the polycondensation of 3-aminophenol and formaldehyde. On the surface of the 3-aminophenol resin microspheres, silver nanoparticles were synthesized in situ and immobilized by simple heating. The composite was employed as a substrate for surface-enhanced Raman scattering (SERS). The SERS enhancement factor was evaluated using 4-mercaptobenzoic acid and Nile blue A as signal molecules. A highly sensitive SERS immunoassay that combined labeled antibody conjugated silver nanoparticle modified 3-aminophenol resin microspheres and coating antibody conjugated magnetic nanoparticles was fabricated to determine carcinoembryonic antigen. A linear relationship was obtained between the Raman intensity and the concentration of carcinoembryonic antigen. The limit of detection was 1.2 picograms per milliliter at a signal-to-noise ratio of three. This is believed to be the first report of a SERS immunoassay using silver nanoparticle modified 3-aminophenol resin microspheres as substrates.     

Synthesis of highly ordered AgNPs-coated silica photonic crystal beads for sensitive and reproducible 3D SERS substrates

3 D highly ordered silver nanoparticles(AgNPs) coated silica photonic crystal beads(Ag/SPCBs) were prepared and exploited as a novel surface enhanced Raman scattering(SERS) substrate.The monodisperse and size-controlled SPCBs were prepared via self-assembly of silica nanoparticles process using a simple microfluidic device.Then the Ag/SPCBs were easily obtained by in situ growth of AgNPs onto the NH₂-modified SPCBs.Field emitting scanning electron microscopy(SEM) and energy dispersive X-ray spectrometry(EDX) were used to characterize the Ag/SPCBs.The effect of silica nanoparticle size and AgNO₃ concentration on the SERS performance of the resultant Ag/SPCBs substrate were discussed in detail.The results indicate that the Ag/SPCBs have highest SERS signals when silica nanoparticle size is250 nm and AgNO₃ concentration is 0.8 mg/mL.Using malachite green(MG) as model analyte,the Ag/SPCBs substrate displayed a high sensitivity and a wide linear range for MG.The well-designed Ag/SPCBs show high uniformity and excellent reproducibility,and can be used as an effective SERS substrate for sensitive assay application.     

Ag纳米粒子在TiO2四棱柱阵列上的可控生长及其SERS效应

采用水热法在导电玻璃FTO导电面上沉积TiO₂四棱柱阵列; 并以其为基体, 分别采用聚乙烯基吡咯 烷酮(PVP)还原Tollens试剂以及柠檬酸三钠(TSC)还原硝酸银溶液, 将Ag纳米粒子(AgNPs)沉积在TiO₂四棱柱阵列上形成TiO₂@AgNPs-PVP和TiO₂@AgNPs-TSC微纳结构作为表面增强拉曼散射(SERS)基底. 实验结果表明, Ag纳米粒子在TiO₂四棱柱阵列上的尺寸和分布可通过改变Tollens试剂的浓度和TSC还原硝酸银溶液的反应时间来调控, 进而优化基底的SERS灵敏度. TiO₂@AgNPs-PVP微纳结构对罗丹明6G(R6G)的检出限为10^-12 mol/L, 对低活性小分子三聚氰胺的检出限为0.01 mg/mL; TiO₂@AgNPs-TSC微纳结构对R6G的检出限为10^-10 mol/L, 对三聚氰胺的检出限为0.01 mg/mL. TiO₂@AgNPs-PVP和TiO₂@AgNPs-TSC微纳结构基底的SERS活性、 循环可回收性与还原剂种类紧密相关: 包覆在Ag纳米粒子上的PVP可以作为隔离层避免Ag纳米粒子直接接触, 防止电磁场耦合作用减弱, 增强基底的SERS活性; 同时, PVP是一种水性聚合物, 有较强的亲水性, 作为循环可回收SERS基底使用时, 吸附小分子物质清洗难度较大.