Disposable, stable media for reproducible surface-enhanced Raman spectroscopy

Large numbers of identical and stable SE(R)RS [surface-enhanced (resonance) Raman]-active media, which are convenient to handle and manipulate but sufficiently inexpensive that they can be used once and then discarded, have been prepared by isolating nanoparticles from Ag and Au sols in hydrophilic polymer gels. The preparation simply involves mixing a suitable polymer with the sol to give a viscous suspension that can be coated onto a substrate and dried to form a hard translucent film. The films remain inactive until they are treated with aqueous analyte solution, which causes the film to swell and brings the analyte into contact with the active metal particles. The swollen films give strong SERS spectra which are effectively identical to those obtained from simple sols. The advantage of this method is that the dried polymers can be stored indefinitely before use and that they give a high degree of spectral reproducibility.     

Metastable state nanoparticle-enhanced Raman spectroscopy for highly sensitive detection

Metastable state silver nanoparticle surface-enhanced Raman scattering has been experimentally and theoretically demonstrated; the signal is two to three orders of magnitude higher than that for the traditional method. Ultrasensitive surface-enhanced Raman scattering signals of illicit drug cocaine and organophosphate pesticide methyl-parathion were observed.     

A highly sensitive detection platform based on surface-enhanced Raman scattering for Escherichia coli enumeration

A very sensitive and highly specific heterogeneous immunoassay system, based on surface-enhanced Raman scattering (SERS) and gold nanoparticles, was developed for the detection of bacteria and other pathogens. Two different types of gold nanoparticles (citrate-stabilized gold nanosphere and hexadecyltrimethylammonium bromide (CTAB)-stabilized gold nanorod particles) were examined and this immunoassay was applied for the detection of Escherichia coli. Raman labels were constructed by using these spherical and rod-shaped gold nanoparticles which were first coated with 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) and subsequently with a molecular recognizer. The working curve was obtained by plotting the intensity of the SERS signal of the symmetric NO₂ stretching of DTNB at 1,333 cm⁻¹ versus the concentration of the E. coli. The analytical performance of gold particles was evaluated via a sandwich immunoassay, and linear calibration graphs were obtained in the E. coli concentration range of 10¹–10⁵ cfu/mL with a 60-s accumulation time. The sensitivity of the Raman label fabricated with gold nanorods was more than three times higher than spherical gold nanoparticles. The selectivity of the developed sensor was examined with Enterobacter aerogenes and Enterobacter dissolvens, which did not produce any significant response. The usefulness of the developed immunoassay to detect E. coli in real water samples was also demonstrated.     

Filter-based surface-enhanced Raman spectroscopy for rapid and sensitive detection of the fungicide ferbam in water

Surface-enhanced Raman spectroscopy (SERS) has been widely applied for rapid and sensitive detection of various chemical and biological targets. Here, we incorporated a filter syringe system into the SERS method to detect the fungicide ferbam in water. Silver nanoparticles (Ag NPs) were aggregated by sodium chloride (NaCl) to form nanoclusters that could be trapped in the pores of the filter membrane to from the SERS-active membrane. Then samples were filtered through the membrane. After capturing the target, the membrane was taken out and air dried before measuring by a Raman instrument. After optimisation of various parameters, the developed filter SERS method was able to detect the fungicide ferbam as low as 2.5 μg/L and had a good quantitative capability. The developed method was successfully applied in three water samples, including double-distilled water, tap water, and pond water. The test can be carried out on site using a portable Raman instrument. This study shows that the filter-based SERS method improves the detection capability in water samples, including the sensitivity and portability, and could be applied in the detection of various toxins in real-world water samples.     

Bioaerosol detection and characterization by surface-enhanced Raman spectroscopy

Instrumentation has been developed to detect and characterize airborne pollen and bacteria rapidly by injecting a bioaerosol into a nanocolloidal suspension of silver particles using a micropump. The biological particles were mixed with the silver colloid in order to deposit the metallic particles on the surface of the bioanalyte. The silver/bioanalyte suspension was pumped through a light scattering cuvette, and the enhanced Raman spectrum was recorded. Surface-enhanced Raman spectra are presented for tree pollen (cottonwood and redwood pollen) and a bacterium (Escherichia coli), and the E. coli spectra are compared with results obtained from the literature and with results obtained previously by mixing various concentrations of the bioanalyte with the silver colloid. Although the system has not been optimized to maximize the Raman spectra, it is shown spectra can be obtained rapidly. Some assignments of the chemical bonds associated with the spectra are based on previously published results for bacteria and pollen.     

Molecularly imprinted polymers for highly sensitive detection of morphine using surface plasmon resonance spectroscopy

Molecular imprinting technology is applied in surface plasmon resonance spectroscopy for highly sensitive and selective detection of morphine（MO）.As SPR-based sensor of MO,the preparation of molecular imprinted polymer is as follows: methacryhc acids（MAA）,ethylene glycol dimethacrylate（EGDMA）,azodiisobutyronitrile（AIBN） were used as functional monomer,cross-linker and initiator,respectively.The experiment results showed that morphine imprinted polymer had the performance of high sensitivity and specificity,i.e.the relative signal of SPR response was proportional to the concentration of morphine in acetonitrile in the range of 10^-9mol/L to 10^-6mol/L（1 ppb-1 ppm） with LOD of 10^-10mol/L,and MO was distinguished from its analogs,such as codeine.     

One-step preparation of porous copper nanowires electrode for highly sensitive and stable amperometric detection of glyphosate

A simple and sensitive amperometric method for the detection of glyphosate based on a porous copper nanowire electrode is presented. The porous nanowires were fabricated simply by a one-step electrodeposition using hydrogen bubbles generated during the deposition to produce pores. These porous nanowires provided a 1.4 times larger surface area than a porous copper film. After the fabrication process the porous copper nanowires can be applied directly as a working electrode with the same custom-built flow cell. The detection was carried out by measuring the oxidation signal of copper; this increased with the concentration of glyphosate due to dissolution of the copper from the electrode. Under optimal conditions, the responses of the sensor were linear between 0.010–5.0 µmiol L^?1, with a limit of detection of 10.0 nmol L^?1 (S/N = 3). The large surface area of the electrode minimised the corrosion effect, as observed by the remarkably stable response (95 injections of 0.20 µmol L^?1 of glyphosate were possible). When applied in order to detect glyphosate in fresh fruit and vegetable samples, the concentrations were found in the range of non-detectable to (0.104 ± 0.005) µmiol L^?1. These results indicated that the fabrication process can be used to produce a new form of working electrode for glyphosate detection.     

Improved MALDI‐MS method for the highly sensitive and reproducible detection of biomarker peaks for the proteotyping of Salmonella serotypes

The rapid identification and classification of pathogenic microorganisms, including Salmonella enterica, is important for the surveillance and prevention of foodborne diseases. Matrix‐assisted laser desorptionionization time‐of‐flight mass spectrometry (MALDI‐TOFMS) has been shown to be an effective tool for the rapid identification of microorganisms. In a previous report, a mass database consisting of 12 biomarker proteins, S8, L15, L17, L21, L25, S7, superoxide dismutase (SodA), peptidylprolyl cis‐trans isomerase C, Gns, YibT, YaiA, and YciF, was introduced for the serotyping of S. enterica via MALDI‐MS (Applied Microbiology and Biotechnology, 2017, 101, 8557‐8569). However, the reproducibility of peak detection of biomarkers such as SodA at mz 23 000 was poor. We report here an optimized MALDI‐MS method for detecting these biomarkers with high sensitivity and reproducibility. The issue was solved by controlling the bacterial concentration at 1 × 10 to 1 × 10² MFU (3 × 10⁶ to 3 × 10⁷ CFUμL, as calculated from the MFU), using the colony suspension supernatant obtained by centrifugation, and using matrix additives such as methylenediphosphonic acid and N‐decyl‐β‐D‐maltopyranoside. We propose that the method including the above steps is one of the best for detecting biomarkers with high sensitivity and reproducibility.     

Solid-liquid-liquid extraction as an approach to the sensitive detection of a hydrophobic pollutant through surface-enhanced Raman spectroscopy

Surface-enhanced Raman scattering (SERS) spectra of thiram (tetramethylthiuram disulfide), a dimethyl dithiocarbamate fungicide, were recorded after the adsorption on plasmonic silver nanowires from a system of water, organic solvent and nanoparticles. As organic solvents dichloromethane and 1-octanol were involved. A method for measuring the adsorption constant of thiram as a model molecule to the silver surface by studying its partition phenomena in a binary solvent system is presented. The method is based on the extraction of a hydrophobic molecule from an organic solvent by an aqueous suspension of silver anisotropic nanoparticles. The obtained results demonstrate the effectiveness of SERS methodology for the sensitive analysis of compounds with low aqueous solubility, and a reliable SERS spectrum of thiram was obtained with excellent signal/noise ratio at low concentrations. In addition, for vibrational assignments, Density Functional Theory (DFT) was used for the simulation of the Raman and SERS spectra of thiram and its complexes with silver considering the following two models: a single silver atom and an Ag₂₀ cluster.     

Preparation of silica-encapsulated hollow gold nanosphere tags using layer-by-layer method for multiplex surface-enhanced raman scattering detection

The use of silica shells offers many advantages in surface-enhanced Raman scattering (SERS)-based biological sensing applications due to their optical transparency, remarkable stability in environmental media, and improved biocompatibility. Here, we report a novel layer-by-layer method for the preparation of silica-hollow gold nanosphere (HGN) SERS tags. Poly(acrylic acid) was used to stabilize Raman reporter-tagged HGNs prior to the adsorption of a coupling agent, after which a silica shell was deposited onto the particle surface using Sto?ber's method. Importantly, competitive adsorption of the Raman reporter molecules and coupling agents, which results in unbalanced loading of reporter molecules on individual nanoparticles, was avoided using this method. As a result, the loading density of reporter molecules could be maximized. In addition, HGNs exhibited strong enhancement effects from the individual particles because of their ability to localize the surface electromagnetic fields through pinholes in the hollow particle structures. The proposed layer-by-layer silica-encapsulated HGN tags showed strong SERS signals as well as excellent multiplexing capabilities.     

A label-free and self-assembled electrochemical biosensor for highly sensitive detection of cyclic diguanylate monophosphate (c-di-GMP) based on RNA riboswitch

Cyclic diguanylate monophosphate (c-di-GMP) is an important second messenger that regulates a variety of complex physiological processes involved in motility, virulence, biofilm formation and cell cycle progression in several bacteria. Herein we report a simple label-free and self-assembled RNA riboswitch-based biosensor for sensitive and selective detection of c-di-GMP. The detectable concentration range of c-di-GMP is from 50 nM to 1 μM with a detection limit of 50 nM.     

A highly stable and sensitive chemically modified screen-printed electrode for sulfide analysis

We report here a highly stable and sensitive chemically modified screen-printed carbon electrode (CMSPE) for sulfide analysis. The CMSPE was prepared by first ion-exchanging ferricyanide into a Tosflex anion-exchange polymer and then sealing with a tetraethyl orthosilicate sol-gel layer. The sol-gel overlayer coating was crucial to stabilize the electron mediator (i.e., Fe(CN)₆³⁻) from leaching. The strong interaction between the oxy-hydroxy functional group of sol-gel and the hydrophilic sites of Tosflex makes the composite highly rigid to trap the ferricyanide mediator. An obvious electrocatalytic sulfide oxidation current signal at ∼0.20 V versus Ag/AgCl in pH 7 phosphate buffer solution was observed at the CMSPE. A linear calibration plot over a wide range of 0.1 μM to 1 mM with a slope of 5.6 nA/μM was obtained by flow injection analysis. The detection limit (S/N = 3) was 8.9 nM (i.e., 25.6 ppt). Practical utility of the system was applied to the determination of sulfide trapped from cigarette smoke and sulfide content in hot spring water.     

HistoFlex--a microfluidic device providing uniform flow conditions enabling highly sensitive, reproducible and quantitative in situ hybridizations

A microfluidic device (the HistoFlex) designed to perform and monitor molecular biological assays under dynamic flow conditions on microscope slide-substrates, with special emphasis on analyzing histological tissue sections, is presented. Microscope slides were reversibly sealed onto a cast polydimethylsiloxane (PDMS) insert, patterned with distribution channels and reaction chambers. Topology optimization was used to design reaction chambers with uniform flow conditions. The HistoFlex provided uniform hybridization conditions, across the reaction chamber, as determined by hybridization to microscope slides of spotted DNA microarrays when applying probe concentrations generally used in in situ hybridization (ISH) assays. The HistoFlex's novel ability in online monitoring of an in situ hybridization assay was demonstrated using direct fluorescent detection of hybridization to 18S rRNA. Tissue sections were not visually damaged during assaying, which enabled adapting a complete ISH assay for detection of microRNAs (miRNA). The effects of flow based incubations on hybridization, antibody incubation and Tyramide Signal Amplification (TSA) steps were investigated upon adapting the ISH assay for performing in the HistoFlex. The hybridization step was significantly enhanced using flow based incubations due to improved hybridization efficiency. The HistoFlex device enabled a fast miRNA ISH assay (3 hours) which provided higher hybridization signal intensity compared to using conventional techniques (5 h 40 min). We further demonstrate that the improved hybridization efficiency using the HistoFlex permits more complex assays e.g. those comprising sequential hybridization and detection of two miRNAs to be performed with significantly increased sensitivity. The HistoFlex provides a new histological analysis platform that will allow multiple and sequential assays to be performed under their individual optimum assay conditions. Images can subsequently be recorded either in combination or sequentially through the ability of the HistoFlex to monitor assays without disassembly.     

Study on self-assembled monolayers of functionalized azobenzene thiols on gold: XPS,electrochemical properties,and surface-enhanced Raman spectroscopy

Self-assembled monolayers (SAMs) of 4-acetamino-4'-(4-mercaptobutoxy)azobenzene (CH3)CONH-ph-N=N-ph-O(CH2)(4)SH, abbr. aaAzoC4SH) and 4-mercaptobutoxy azobenzene (ph-N=N-ph-O(CH2)(4)SH, abbr. AzoC4SH) on a gold surface have been studied by X-ray photoelectron spectroscopy (XPS), FT Raman spectroscopy, and electrochemistry. A surface-enhanced Raman scattering (SERS)-active system with a "sandwiched" structure of Ag/R-Azo-C4S-/Au was conveniently obtained by the method of Tollen's test. The relationship between the SERS effect and the structural nature of the system indicates that the enhancement correlates to both the silver islands above and the gold substrate underneath. The redox behaviors of the self-assembly on gold electrodes showed that the SAMs of the two compounds exhibit well-behaved voltammetric responses in a Britton-Robinson buffer corresponding to the irreversible two-electron, two-proton reduction-oxidation of azobenzene. The apparent electron-transfer rate kinetics is very sluggish, and the rate constant k(app) of aaAzoC4SH/Au (1.34 x 10(-6) s(-1)) is lower than that of AzoC4SH/Au (1.63 x 10(-4) s(-1)), which may be attributed to the different spatial restriction of close-packing structures on the conformational change accompanied by electron and proton transfer in the SAMs.     

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).     

Rich-defect carbon nanotubes for highly sensitive detection of Dopamine

Dopamine (DA) plays an essential role in the central nervous, renal, hormonal and cardiovascular systems. Various modified carbon nanotubes (CNT)-based dopamine sensors have been reported, but inexpensive, highly sensitive plain CNT-based ones are seldom studied. In this work, a facile and inexpensive CNT-based DA sensor is made by rich-defect multi-walled carbon nanotubes (RD-CNT) via an ultrasound method. The defect and elemental states of the RD-CNT are systematically studied by transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HR-TEM), Raman spectroscopy, X-ray powder diffraction (XRD) and X-ray-photoelectron spectroscopy (XPS). Results show that massive holes and cracks exist in RD-CNT. The level of defects increases from the additional exposed edges. The electrochemical characterizations indicate that the electrochemical sensor has the highest sensitivity of 438.4 μA/(μM ⋅ cm²) among all carbon materials-based DA sensors while well meeting the clinically required detection range and selectivity. The DA sensor was further used to detect live healthy human serum and live PC12 cells with satisfactory results, thus holding great promise for an inexpensive but sensitive DA sensor in practical applications of clinical diagnosis and biological research.     

A self-assembled deoxyribonucleic acid concatemer for sensitive detection of single nucleotide polymorphism

Polymerase-free and label-free strategies for DNA detection have shown excellent sensitivity and specificity in various biological samples. Herein, we propose a method for single nucleotide polymorphism (SNP) detection by using self-assembled DNA concatemers. Capture probes, bound to magnetic beads, can joint mediator probes by T4 DNA ligase in the presence of target DNA that is complementary to the capture probe and mediator probe. The mediator probes trigger self-assembly of two auxiliary probes on magnetic beads to form DNA concatemers. Separated by a magnetic rack, the double-stranded concatemers on beads can recruit a great amount of SYBR Green I and eventually result in amplified fluorescent signals. In comparison with reported methods for SNP detection, the concatemer-based approach has significant advantages of low background, simplicity, and ultrasensitivity, making it as a convenient platform for clinical applications. As a proof of concept, BRAF^T1799A oncogene mutation, a SNP involved in diverse human cancers, was used as a model target. The developed approach using a fluorescent intercalator can detect as low as 0.1 fM target BRAF^T1799A DNA, which is better than those previously published methods for SNP detection. This method is robust and can be used directly to measure the BRAF^T1799A DNA in complex human serum with excellent recovery (94–103%). It is expected that this assay principle can be directed toward other SNP genes by simply changing the mediator probe and auxiliary probes.     

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.