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A feedback inhibition effect of high autoinducer levels on metabolite secretion in Chromobacterium subtsugae (C. subtsugae) was evidenced by in situ spatiotemporal surface-enhanced Raman spectroscopy (SERS) profiling. The hierarchical hydrophobic plasmonic array in agar medium is structured by oil/water/oil (OL/W/OH) triphasic interfacial self-assembly. The hydrophobic layer acts as a "door curtain" to selectively permit adsorption of a quorum sensing (QS)-regulated fat-soluble metabolite, i.e., violacein (Vio), and significantly blocks nonspecific adsorption of water-soluble proteins, etc. The SERS profiling clearly evidences that the diffusion of N-hexanoyl-l-homoserine lactone (C6-HSL) in agar medium quickly triggers the initial synthesis of Vio in C. subtsugae CV026 but surprisingly inhibits the intrinsic synthesis of Vio in C. subtsugae ATCC31532. The latter negative response might be related to the VioS repressor of ATCC31532, which negatively controls violacein production without influencing the expression of the CviI/R QS system. Moreover, two sender-receiver systems are constructed by separately coculturing CV026 or ATCC31532 with Hafnia alvei H4 that secretes large amounts of C6-HSL. Expectedly, the cocultivation similarly triggers the initial synthesis of Vio in CV026 but seems to have a quite weak negative effect on the intrinsic synthesis in ATCC31532. In fact, the negative regulation in ATCC31532 might be affected by a diffusion-dependent concentration effect. The H4 growth and its secretion of C6-HSL are a slow and continuous process, thereby avoiding the gathering of local high concentrations. Overall, our study put forward an in situ SERS strategy as an alternative to traditional bioluminescent tools for highly sensitively analyzing the spatiotemporal communication and cooperation in live microbial colonies.
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Bacterias , Percepción de Quorum , Agar , Chromobacterium/fisiologíaRESUMEN
An efficient method for the synthesis of alkynyl sulfides via a C(sp3)-S bond cleavage of α-bromostyrene sulfonium salts has been developed. This base-promoted nucleophilic ring-opening pathway allows the preparation of diverse alkynyl sulfide compounds using tetramethylene sulfoxide as the sulfur source. The reaction proceeds with good functional group tolerance and could be applied to the late-stage functionalization of bioactive molecules and drugs. Furthermore, the synthetic utility of this method was demonstrated by a one-pot synthesis, scale-up reaction and further modification of various alkynyl sulfide products.
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Controllably regulating the electrostatic bilayer of nanogold colloids is a significant premise for synthesizing spherical nucleic acid (SNA) and building ordered plasmonic architectures. We develop a facile acoustic levitation reactor to universally synthesize SNAs with an ultra-high density of DNA strands, which is even higher than those of various state-of-the-art methods. Results reveal a new mechanism of DNA grafting via acoustic wave that can reconfigure the ligands on colloidal surfaces. The acoustic levitation reactor enables substrate-free three-dimentional (3D) spatial assembly of SNAs with controllable interparticle nanogaps through regulating DNA lengths. This kind of architecture may overcome the plasmonic enhancement limits by blocking electron tunneling and breaking electrostatic shielding in dried aggregations. Finite element simulations support the architecture with 3D spatial plasmonic hotspot matrix, and its ultrahigh surface-enhanced Raman scattering (SERS) capability is evidenced by in situ untargeted tracking of biomolecular events during photothermal stimulation (PTS)-induced cell death process. For biomarker diagnosis, the conjugation of adenosine triphosphate (ATP) aptamer onto SNAs enables in situ targeted tracking of ATP during PTS-induced cell death process. Particularly, the CD71 receptor and integrin α3ß1 protein on PL45â cell membrance could be well distinguished by label-free SERS fingerprints when using specific XQ-2d and DML-7 aptamers, respectively, to synthesize SNA architectures. Our current acoustic levitation reactor offers a new method for synthesizing SNAs and enables both targeted and untargeted SERS analysis for tracking molecular events in living systems. It promises great potentials in biochemical synthesis and sensing in future.
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Oro , Espectrometría Raman , Oro/química , ADN/química , Nanopartículas del Metal/química , Acústica , Humanos , Propiedades de Superficie , Adenosina Trifosfato/químicaRESUMEN
Surface-enhanced Raman spectroscopy (SERS) with ultrasensitive vibrational fingerprints enables quick identification and trace detection of various kinds of molecules. But proteins usually have low Raman cross sections and are difficult to generate recognizable signals in direct SERS detection. Recently, nucleic acid-peptide conjugates are emerging with great potential in structuring, assembling, catalyzing, sensing, etc., and the coupling of aptamers further enables superior biological recognition and programmability. Here, we develop the aptamer-peptide conjugates as a new kind of SERS probe for direct high-specific profiling abnormal protein levels in cancer patients. The aptamer conjugated with glutathione (GSH) functions as both the recognition element and the SERS reporters that can simultaneously generate SERS fingerprints of both peptides and nucleic acids. This kind of biocompatible probe appears to have excellent performance in high-salt environments and realizes rapid, simple, and multisignal detection of thrombin (TB). Data-driven soft independent modeling of class analogy (DD-SIMCA) is used to distinguish SERS profiles of actual blood samples and realize the identification and classification of cancer patients. Furthermore, the effect of low-temperature storage time on blood samples is analyzed by tracking the changes of SERS profiles; the results hint that plasma samples stored under 4 °C for more than 2 days could generate false negative results due to TB hydrolysis, which has important implications for clinical sample analysis. This kind of nucleic acid-peptide conjugate provides new ideas for SERS sensing strategy in the future.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Nanopartículas del Metal , Neoplasias , Ácidos Nucleicos , Humanos , Espectrometría Raman/métodos , Proteínas , Péptidos , Aptámeros de Nucleótidos/química , Neoplasias/diagnóstico , Nanopartículas del Metal/química , Oro/química , Técnicas Biosensibles/métodosRESUMEN
The rational design of ice recrystallization inhibition (IRI) materials is challenging due to the poor understanding of the IRI mechanism at the molecular level. Here we report several new findings about IRI. (1) A dense hydroxyl monolayer of small molecules, e.g. 6-aza-2-thiothymine (ATT), adsorbed on a nanogold surface was demonstrated, for the first time, to have IRI activity. Five structural analogues adsorbed on groups nanogold with outward hydroxyl or methyl were created to evidence the origin of IRI activity. (2) Their IRI mechanism is closely related to the density of hydroxyls on a nanogold surface. However, the hydrophobic interaction in our model is not essential for macroscopic IRI activity. (3) A molecular dynamics simulation elucidates the hydroxyl density dependent IRI trajectories underlying the experimental observations, and the radial distribution function reveals that the methyl even slightly hinders the formation of hydrogen bonding due to a hydrophobic interaction. This work sheds more light on the IRI mechanism that should help in the customization of novel IRI materials.
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Hielo , Simulación de Dinámica Molecular , Cristalización , Interacciones Hidrofóbicas e HidrofílicasRESUMEN
The lipids with a rich diversity of isomers face a formidable challenge in comprehensive structural analysis. The commonly used mass spectrometry-based techniques usually require a considerable number of molecules with sophisticated chemical derivatization or ion mobility separation, but the co-existing of structurally similar isomers often makes the distinction impossible. Here, we develop an alternative powerful liquid/liquid interfacial surface-enhanced Raman spectroscopy (SERS) strategy at normal temperature and pressure without any sources of ionization or radiation. This strategy generates high-resolution fingerprints in molecular chain length, CâC position, saturation, and regio- and stereoisomers of both glycerides and fatty acids and requires only trace amounts of molecules down to 1 ppb to achieve discrimination and exhibits great potentials to push the identification capability to trace levels or even the single-molecule level. According to experimental data and theoretical simulations, these targets have the amphiphilic and emulsifying properties, exhibit ordered molecular orientation and adsorption patterns, promote the co-assembly with plasmonic nanoarrays at the immiscible liquid/liquid interface, and consequently amplify the detection sensitivity. As a contrast, the typical SERS based on solid/air interfacial plasmonic nanoarrays faces the intrinsic bottleneck of extremely weak intensity and indistinguishable spectral fingerprints of lipid molecules. The vibrational fingerprints exhibit a rich range of well-resolved absorption features that are clearly diagnostic for fine structural changes and pave a new way for straightforward measurement without laborsome sample purification, enrichment, or complex derivatization. Although challenging, its unprecedented resolving power expands the potentials of SERS, serving as an ultimate analytical method to provide insights into the detailed structural features of other lipids under facile conditions in the future.
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Alcanos , Espectrometría Raman , Adsorción , Espectrometría de Masas , Espectrometría Raman/métodos , EstereoisomerismoRESUMEN
High-throughput surface-enhanced Raman scattering (SERS) reader, especially for liquid sample testing, is of great significance and huge demand in biology, environment, and other analytical fields. Inspired by the principle of microplate reader, herein, we developed a microplate-SERS reader for semiautomatic and high-throughput assays by virtue of three-dimensional liquid interfacial arrays (LIAs). For the first time, the formation of LIA in oil-in-water state, water-in-oil state, and two-dimensional plane state is realized by operating the hydrophilicity (contact angle) of the container. Through the force analysis of LIA, the effect of organic (O) phase density on the relative position of LIA was quantified. In addition, the optimized reader offers fast and continuous semiautomatic detection of 12 samples below 10 min with great signal reproducibility (calibration with the characteristic peak of O phase as the internal standard). The isolated wells in the microplate prevent analyte cross talk, allowing accurate quantification of each sample. Multiplex analysis capability highlights that this reader has the ability of rapid identification and quantification of samples containing various analytes and concentrations. The results demonstrate high-resolution dual and triple analyte detection with fully preserved signal and Raman features of individual analytes in a mixture, which implies that it also has excellent anticounterfeiting applications. This microplate-SERS reader combines the superior advantages of the LIA, microplate, and SERS techniques to retrieve the molecular vibrational fingerprints of various chemicals in complex media.
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Espectrometría Raman , Agua , Interacciones Hidrofóbicas e Hidrofílicas , Reproducibilidad de los Resultados , Espectrometría Raman/métodosRESUMEN
For surface-enhanced Raman scattering (SERS) analysis, only analytes that can be absorbed spontaneously onto a noble metal surface can be detected effectively. Therefore, getting nonadsorptive molecules close enough to the surface has always been a key challenge in SERS analysis. Here absorbance measurements show that the liquid-interfacial array (LIA) does not adsorb or enrich benzopyrene (Bap) molecules, which lack effective functional groups that can interact with the noble metal surfaces. But the SERS intensity of 0.1 ppm Bap on the LIA is 10 times larger than that of 10 ppm Bap on traditional solid substrate, i.e., 3 orders of magnitude of enhancement. The LIA overcomes the restriction of affinity between Bap molecules and the metal surface, and the Bap molecules can easily enter nanogaps without steric hindrance. Furthermore, both adsorptive and nonadsorptive molecules were used to observe the SERS enhancement behavior on the LIA platforms. In multiple detection, competitive SERS signal changes could be observed between adsorptive and nonadsorptive molecules or between nonadsorptive and nonadsorptive molecules. A theoretical scheme was profiled for localized surface plasmon resonance (SPR) properties of the LIA. Finite difference-time domain (FDTD) simulation shows that the LIAs have biphasic and accessible asymmetric hotspots, and the electric field enhancement in the CHCl3 (O) phase is approximately four times larger than that of the water (W) phase. In addition, the position and relative strength of the electromagnetic field depend on the spatial position of gold nanoparticles (GNPs) relative to the liquid-liquid interface (LLI), i.e., when the GNP dimer is completely immersed in a certain phase, the electromagnetic field enhancement of the CHCl3 phase is approximately 7 times larger than that of the W phase. We speculate that dual-phase-accessible hotspots and the hydrophobic environment provided by CHCl3 are two important factors contributing to successful detection of four common polycyclic aromatic hydrocarbons (PAHs) with a detection limit of 10 ppb. Finally, the LIA platform successfully realizes simultaneous detection of multiple PAHs in both plant and animal oils with good stability. This study provides a new direction for the development of high-efficiency and practical SERS technology for nonadsorptive molecules.
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Liquid-state interfacial plasmonic systems are emerging as an alternative for the quantitation and practicability of the surface-enhanced Raman scattering (SERS) technique in analytical science, especially for complex liquid-phase systems. Here we show a general strategy for the three-dimensional (3D) self-assembly of gold nanoparticle (GNP) arrays on a spherical oil-water (O-W) interface, denoted as a plasmonic metal liquid (PML). The PML has excellent self-healing and shape-adaptive features; it can be transferred into containers of any shape; and it presents fast, quantitative, and multiplex SERS capability. Accurate control of nanoparticle density (PD) on the 3D interface enables tunable SERS strength. In situ synchrotron radiation small angle X-ray scattering (SR-SAXS) provides evidence that the interfacial PD is quantifiable and can be precisely regulated in the range of 24-216 particles/µm2, which produces optimizable Raman enhancement. The strongest SERS signal is achieved at 167 particles/µm2 with GNP diameters of approximately 64 nm. In particular, the O phase acts not only as the assembly media for spherical PML arrays but also as the extracting agent for targets with different natures in complex media. Moreover, the O phase with continuous-phase features generates inherent and sharp SERS fingerprints and provides an effective internal standard (IS) for calibrating the fluctuation of samples and measuring conditions. By virtue of the triple roles of the O phase, the PML platform exhibits excellent mechanical stability, detection sensitivity, and signal reproducibility. This study demonstrates the concept of a fast and quantitative liquid-state SERS platform in common cuvettes on a portable Raman device that is as simple as a spectrophotometer.
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Liquid-state interfacial nanoparticle arrays for surface-enhanced Raman scattering (SERS) promises a practical, substrate-free, and rapid analysis but faces a great challenge to develop a batch and uniform fabrication strategy with stable internal standards (IS) because of the difficulties in precisely locating both the IS tags and analytes in the same local structure under the harsh conditions of biphasic liquid interface. Here, we develop a fast batch preparation of self-ordered dense Au nanoparticle (GNP) arrays on cyclohexane/water biphasic interface in 96-well plates with the assist of acetone as the phase-crossing inducer. The acetone can extract the pesticide molecules via a simple dipping sample peels and can rapidly capture and locate the pesticide molecule into the plasmonic hotspots. Meanwhile, this phase-crossing solvent, acetone itself, generates stable SERS signal and is used as the IS tags to calibrate the signal fluctuation. This platform presents an excellent uniformity with a relative standard deviation (RSD) of 5.9% compared to the RSD of 14.5% without the IS's correction and a good sensitivity with a limit of detection (LOD) of 1 nM thiram. This high-throughput strategy for analyzing pesticide residues at fruit peels reached detection levels of nanograms per square centimeter (ng/cm2). Combined with the 96-well plates, this platform greatly facilitates the self-assembly and multiplex sampling. The self-ordered arrays at two immiscible phases interface evidenced the detection of both the oil-soluble thiabendazole and the water-soluble thiram molecules and also realized the multiplex and two-phase detection of these two pesticides. This platform offers vast possibilities for on-site sensing of various analytes and paves a new way for the quantitative and high-throughput SERS analyzer just as convenient as the microplate reader.
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Poor adsorption properties of nonadsorbing targets and competing adsorption of nontargets at a liquid interface always hamper the development of interface sensing techniques. There is a need to fabricate materials that are applicable to various interface assemblies and, meanwhile, could be employed as interfacial gating to improve the performance of interface sensing by separating, enriching, and recognizing targets at the liquid interface. Here, superhydrophobic zeolite imidazole frameworks-8@gold nanoparticles-1H,1H,2H,2H-perfluorodecanethiol (ZIF-8@GNPs-PFDT) with a static water contact angle (WCA) of 155° was constructed via electrostatic self-assembly and surface graft modification. The plasmonic metal-organic framework (PMOF) nanohybrid realized all-purpose self-assembly at air/liquid and liquid/liquid interfaces and also facilely assembled on the surface of liquid droplets, hydrogels, and foams. The self-assembled porous materials displayed the capability for separating, enriching, and recognizing analytes at various oil/water interfaces and thus could be used to adsorb nonadsorbing targets and block the competing adsorption of nontargets. The self-assembled ZIF-8@GNPs-PFDT structures were employed as a three-in-one interfacial gating to endow the excellent surface-enhanced Raman scattering (SERS) sensing capability and has become a promising tool for dye molecular analysis, oil/water separation, organic phase identification, and in situ cultivation and monitoring of bacterial quorum sensing (QS).
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Oro , Interacciones Hidrofóbicas e Hidrofílicas , Estructuras Metalorgánicas , Percepción de Quorum , Estructuras Metalorgánicas/química , Oro/química , Nanopartículas del Metal/química , Zeolitas/química , Adsorción , Propiedades de SuperficieRESUMEN
Salt stress represents a significant abiotic stressor for plants and poses a severe threat to agricultural productivity. Glutaredoxins (GRXs) are small disulfide reductases that can scavenge cellular reactive oxygen species and are crucial for plant growth and development, particularly under stressful circumstances. Although CGFS-type GRXs were found to be involved in various abiotic stresses, the intrinsic mechanism mediated by LeGRXS14, a tomato (Lycopersicon esculentum Mill.) CGFS-type GRX, is not yet fully understood. We discovered that LeGRXS14 is relatively conserved at the N-terminus and exhibits an increase in expression level under salt and osmotic stress conditions in tomatoes. The expression levels of LeGRXS14 in response to osmotic stress peaked relatively rapidly at 30 min, while the response to salt stress only peaked at 6 h. We constructed LeGRXS14 overexpression Arabidopsis thaliana (OE) lines and confirmed that LeGRXS14 is located on the plasma membrane, nucleus, and chloroplasts. In comparison to the wild-type Col-0 (WT), the OE lines displayed greater sensitivity to salt stress, resulting in a profound inhibition of root growth under the same conditions. Analysis of the mRNA levels of the WT and OE lines revealed that salt stress-related factors, such as ZAT12, SOS3, and NHX6, were downregulated. Based on our research, it can be concluded that LeGRXS14 plays a significant role in plant tolerance to salt. However, our findings also suggest that LeGRXS14 may act as a negative regulator in this process by exacerbating Na+ toxicity and the resulting oxidative stress.
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The adverse effects of recrystallization limit the application of cryopreservation in many fields. Peptide-based materials play an essential role in the antifreezing area because of their excellent biocompatibility and abundant ice-binding sites. Peptide-gold nanoparticle conjugates can effectively reduce time and material costs through metal-thiol interactions, but controlled modification remains an outstanding issue, which makes it difficult to elucidate the antifreezing effects of antifreeze peptides at different densities and lengths. In this study, we developed an instant peptide capping on gold nanoparticles with butanol-assisted dehydration and provided a controllable quantitative coupling within a certain range. This chemical dehydration makes it possible to fabricate peptide-gold nanoparticle conjugates in large batches at minute levels. Based on this, the influence of the peptide density and sequence length on the antifreezing behaviors of the conjugates was investigated. The results evidenced that the antifreezing property of the flexible peptide conjugated on a rigid core is related to both the density and length of the peptide. In a certain range, the density is proportional to the antifreeze, while the length is negatively correlated with it. We proposed a rapidly controllable method for synthesizing peptide-gold nanoparticle conjugates, which may provide a universal approach for the development of subsequent recrystallization-inhibiting materials.
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Cristalización , Oro , Hielo , Nanopartículas del Metal , Deshidratación , Oro/química , PéptidosRESUMEN
The authors wish to correct the following error in the original paper [...].
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In situ monitoring of chemical reactions has attracted great attention in many fields. Herein, we successfully in situ track the degradation reaction process of a dye pollutant, methylene blue (MB), on the liquid-liquid interface (LLI) of bimetallic gold core-silver shell nanoparticles (Au@AgNPs) by surface-enhanced Raman spectroscopy (SERS). The optimized LLI bimetallic array of Au50@Ag10NPs exhibits ultrahigh SERS enhancement and excellent catalytic activity. Results evidenced a detection limit of MB down to 1 ppb, and the degradation rate of Au@AgNPs was as high as 85.2% in 30 s, relying on the excellent self-healing properties of nanoarrays. Furthermore, as a practical SERS analyzer, the LLI bimetallic array was used to detect trace amounts of other harmful dyes, including Rhodamine 6G (R6G) and crystal violet (CV) in pure or complex media. Our LLI bimetallic array exhibits a new orientation for monitoring catalytic reactions involving highly toxic, hazardous, or costly targets in food security fields in the future.
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Toward the challenge on reliable determination of trace fentanyl to avoid opioid overdose death in drug crisis, here we realize rapid and direct detection of trace fentanyl in real human urine without pretreatment by a portable surface enhanced Raman spectroscopy (SERS) strategy on liquid/liquid interfacial (LLI) plasmonic arrays. It was observed that fentanyl could interact with the gold nanoparticles (GNPs) surface, facilitate the LLI self-assembly, and consequently amplify the detection sensitivity with a limit of detection (LOD) as low as 1 ng/mL in aqueous solution and 50 ng/mL spiked in urine. Furthermore, we achieve multiplex blind sample recognition and classification of ultratrace fentanyl doped in other illegal drugs, which has extremely low LODs at mass concentrations of 0.02% (2 ng in 10 µg of heroin), 0.02% (2 ng in 10 µg of ketamine), and 0.1% (10 ng in 10 µg of morphine). A logic circuit of the AND gate was constructed for automatic recognition of illegal drugs with or without fentanyl doping. The data-driven analog soft independent modeling model could quickly distinguish fentanyl-doped samples from illegal drugs with 100% specificity. Molecular dynamics (MD) simulation elucidates the underlying molecular mechanism of nanoarray-molecule co-assembly through strong π-metal interactions and the differences in the SERS signal of various drug molecules. It paves a rapid identification, quantification, and classification strategy for trace fentanyl analysis, indicating broad application prospects in response to the opioid epidemic crisis.
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Drogas Ilícitas , Ketamina , Nanopartículas del Metal , Humanos , Morfina/orina , Heroína/análisis , Fentanilo/orina , Oro/química , Nanopartículas del Metal/química , Espectrometría Raman/métodosRESUMEN
The ubiquity of micro-/nanoplastics poses a visible threat to the environment, aquatic organisms, and human beings and has become a global concern. Here, we proposed a liquid interface-based strategy using surface-enhanced Raman spectroscopy to coassemble nanoplastics and gold nanoparticles into a dense and homogeneous plasmonic array, thereby enabling the rapid and sensitive detection of trace nanoplastics. In addition, due to the uniqueness of the oil-water immiscible two-phase interface, we achieved ideal results for the detection of nanoplastics in a complex matrix (e.g., aqueous environment and edible oil) with a detection limit of µg/mL. With the aid of the principal component analysis algorithm, the differentiation and identification of multiple nanoplastic components (e.g., polystyrene, polyethylene, and polyethylene terephthalate) in aqueous environments and common hazards (e.g., Bap and Phe) in edible oil were achieved. Therefore, our self-assembled plasmonic arrays are expected to be used for monitoring environmental pollution and food safety.
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Nanopartículas del Metal , Microplásticos , Humanos , Nanopartículas del Metal/química , Espectrometría Raman/métodos , Oro/química , Agua/química , AceitesRESUMEN
Highly sensitive and reliable detection of ß-adrenergic agonists is especially necessary due to the illegal abuse of growth-promoting feed additives. Here, we develop a novel surface plasmon resonance/surface-enhanced Raman scattering (SPR/SERS) dual-mode plasmonic sensor based on core-satellite nanoassemblies for the highly sensitive and reliable detection of ractopamine (RAC). The addition of RAC results in the decomposition of core-satellite nanoassemblies and consequently changes the Rayleigh scattering color of dark-field microscopy (DFM) images and the Raman scattering intensity of SERS spectra. The excellent sensitivity, specificity, and uniformity of this strategy were confirmed by detecting RAC in various complex media in the farm-to-table chain, and the limit of detection (LOD) was 0.03 ng/mL in an aqueous solution. In particular, the convenient access to livestock sewage not only ensures animal welfare but also provides great convenience for the market regulation of ß-agonists. The success of our on-site strategy only with a portable Raman device promises great application prospects for ß-agonist detection.
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Nanopartículas del Metal , Resonancia por Plasmón de Superficie , Animales , Resonancia por Plasmón de Superficie/métodos , Oro , Espectrometría Raman/métodosRESUMEN
The kiwi plant is dioecious, and its sex is generally identified from flower morphology at blossoming, which takes several years. It is quite necessary but challenging to on-spot identify the plant sex in juvenile stage. Here the target DNA was obtained by screening the Friendly boy (FrBy) gene which is sex-related for different kiwi plant species. Its complementary sequence was divided into two parts as primer DNA and further attached to different gold nanoparticles (GNPs). The connection between target DNA and primer DNA will promote the formation of plasmonic dimers. Dark field microscopy (DFM) can distinguish particles in different aggregation states. Various conditions were optimized based on the standard of increasing the proportion of dimers while reducing that of large aggregates. Furthermore, two Raman reporters (RR) are separately labeled on the nanoprobes, and the plasmonic dimers lead to a tremendous Raman enhancement of two reporters located at the dimer nanogap. Double-blind tests proved the feasibility of this method on the actual samples of kiwi plant leaves. Our SERS method is sensitive, specific, and reliable for rapid sex identification analysis at the kiwi seeding stage, with great promise for decision-making in field management.
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Actinidia , Nanopartículas del Metal , Humanos , ADN , Oro , Polímeros , Espectrometría Raman/métodos , Método Doble CiegoRESUMEN
In situ identification of aptamer-binding targets on living cell membrane surfaces is of considerable interest, but a major challenge, specifically, when advancing recognition to the level of membrane receptor subunits. Here we propose a novel nanometal surface energy transfer (NSET) based nanoruler with a single-nucleobase resolution (SN-nanoruler), in which FAM-labeled aptamers and single-sized gold nanoparticle (GNP) antibody conjugates act as a donor and an acceptor. A single nucleobase resolution of the SN-nanoruler was experimentally illustrated by molecular size, orientation, quenching nature, and other dye-GNP pairs. The SN-nanoruler provides high reproducibility and precision for measuring molecule distance on living cell membranes at the nanometer level owing to only the use of single-sized antibody-capped GNPs. In situ identification of the aptamer binding site was advanced to the protein subunit level on the living cell membrane for the utilization of this SN-nanoruler. The results suggest that the proposed strategy is a solid step towards the wider application of optical-based rulers to observe the molecular structural configuration and dynamic transitions on the membrane surface of living cells.