Au-Ag Nanoshells Research Articles

Au-Ag nano-garlands as a versatile SERS substrate: Two-step synthesis realizes the growth of petal-shaped branches on hollow Au-Ag nanoshells

In this work, we synthesized Au-Ag alloy nano-garlands with multiple petal-like branches and internal cavities in two steps. First, we etched silver nanospheres with HAuCl4 to obtain Au-Ag alloy nanoshells with different shell layer thicknesses and hollow core sizes. After that, based on different nanoshells, 4-NTP was used as a shape-directing agent, HAuCl4 and AgNO3 as precursor raw materials, and AA as a reducing agent for the growth of petals to form Au-Ag bimetallic nano-garlands with multi-branching structure. The regulation of the branching morphology of the nano-garlands could be achieved by changing the amounts of 4-NTP and HAuCl4. The SERS activity of the nano-garlands could be further improved by changing the AA concentration, the size of nanoshells, and the Au/Ag ratio of the precursor raw materials. Because the bimetallic nano-garlands have hollow cavities and petal-like branches, there are couplings between the cavities and the branches, between the adjacent branches, and between the metals inside and outside of the cores, which can achieve a good enhancement of SERS. The highest SERS enhancement factor of Au-Ag bimetallic nano-garlands can reach 4.47×108, which can be used as a good candidate for SERS substrate for the trace detection of biomolecules, pesticides, and other substances.

Near-infrared II plasmonic porous cubic nanoshells for in vivo noninvasive SERS visualization of sub-millimeter microtumors

In vivo surface-enhanced Raman scattering (SERS) imaging allows non-invasive visualization of tumors for intraoperative guidance and clinical diagnostics. However, the in vivo utility of SERS is greatly hampered by the strong optical scattering and autofluorescence background of biological tissues and the lack of highly active plasmonic nanostructures. Herein, we report a class of porous nanostructures comprising a cubic AuAg alloy nanoshell and numerous nanopores. Such porous nanostructures exhibit excellent near-infrared II plasmonic properties tunable in a broad spectral range by varying the pore features while maintaining a small dimension. We demonstrate their exceptional near-infrared II SERS performance varying with the porous properties. Additionally, near-infrared II SERS probes created with porous cubic AuAg nanoshells are demonstrated with remarkable capability for in vivo visualization of sub-millimeter microtumors in a living mouse model. Our near-infrared II SERS probes hold great potentials for precise demarcation of tumor margins and identification of microscopic tumors.

SERS Immunosensor Based on High-Density “Hotspots” Au@SiO2 Array Substrate and Au-Ag Nanoshells Probes for Ultrasensitive Detection of Dual Biomarkers in the Cervical Cancer Serum

Cervical cancer is a common cause of cancer-related death in women worldwide, and quantitative detection of serum tumor markers has practical implications for diagnosing cervical cancer. In this study, a novel surface-enhanced Raman scattering (SERS)-based immunosensor for the hypersensitive simultaneous detection of survivin and osteopontin (OPN) in cervical cancer serum was proposed, which was composed of the AuNPs@SiO2 microspheres (Au@SiO2) array immunosubstrate and Au-Ag nanoshells (Au-AgNSs) immunoprobes. The experimental results demonstrated that the constructed SERS immunosensor exhibited satisfactory selectivity and reproducibility while possessing detection limits as low as 0.908[Formula: see text]pg/mL and 0.813[Formula: see text]pg/mL for survivin and OPN in human serum. Finally, its usefulness in real serum samples (from healthy subjects, cervical intraepithelial neoplasia 1 (CIN 1), CIN 2, CIN 3 and cervical cancer patients) was verified, and the detection data were in good agreement with the enzyme-linked immunosorbent assay results. This immunosensor has excellent performance, which is promising for the early screening of cancer.

Plasmonic LAMP: Improving the Detection Specificity and Sensitivity for SARS-CoV-2 by Plasmonic Sensing of Isothermally Amplified Nucleic Acids.

The ability to detect pathogens specifically and sensitively is critical to combat infectious diseases outbreaks and pandemics. Colorimetric assays involving loop‐mediated isothermal amplification (LAMP) provide simple readouts yet suffer from the intrinsic non‐template amplification. Herein, a highly specific and sensitive assay relying on plasmonic sensing of LAMP amplicons via DNA hybridization, termed as plasmonic LAMP, is developed for the severe acute respiratory syndrome‐related coronavirus 2 (SARS‐CoV‐2) RNA detection. This work has two important advances. First, gold and silver (Au–Ag) alloy nanoshells are developed as plasmonic sensors that have 4‐times stronger extinction in the visible wavelengths and give a 20‐times lower detection limit for oligonucleotides over Au counterparts. Second, the integrated method allows cutting the complex LAMP amplicons into short repeats that are amendable for hybridization with oligonucleotide‐functionalized Au–Ag nanoshells. In the SARS‐CoV‐2 RNA detection, plasmonic LAMP takes ≈75 min assay time, achieves a detection limit of 10 copies per reaction, and eliminates the contamination from non‐template amplification. It also shows better detection specificity and sensitivity over commercially available LAMP kits due to the additional sequence identification. This work opens a new route for LAMP amplicon detection and provides a method for virus testing at its early representation.

A sandwich SERS immunoassay platform based on a single-layer Au-Ag nanobox array substrate for simultaneous detection of SCCA and survivin in serum of patients with cervical lesions.

The evaluation of tumor biomarkers in blood specimens is vital for patients with cervical lesions. Herein, an ultrasensitive surface enhanced Raman scattering (SERS) platform was proposed for simultaneous detection of cervical-lesion-related serum biomarkers. Raman reporter labeled Au–Ag nanoshells (Au–AgNSs) acted as SERS tags and an Au–Ag nanobox (Au–AgNB) array substrate prepared by the oil–water interface self-assembly method was used as a capture substrate. This single-layer Au–AgNB array substrate was proved to have exceptional uniformity by atomic force microscopy and SERS mapping. Numerous “hot spots” and specific adsorption surfaces offered by the Au–AgNB array substrate were confirmed by the finite difference time domain method, which could generate a SERS signal in electromagnetic enhancement. Binding of antigens between antibodies on Au–AgNSs and the Au–AgNB array substrate led to the formation of a sandwich-structure by the two metal nanostructures. Consequently, an ultralow detection limit of 6 pg mL−1 for squamous cell carcinoma antigen (SCCA) and 5 pg mL−1 for survivin in a wide linear logarithmic range of 10 pg mL−1 to 10 μg mL−1 was acquired. High selectivity and reproducibility with relative standard deviations of 7.701% and 6.943% were detected. Furthermore, the simultaneous detection of the two biomarkers in practical specimens was conducted, and the results were consistent with those of the enzyme-linked immunosorbent assay. This platform exhibited good robustness in the rapid and sensitive detection of SCCA and survivin, which could be a promising tool in early clinical diagnosis for different grades of cervical lesions.

Detection of resistance protein A (MxA) in paper-based immunoassays with surface enhanced Raman spectroscopy with AuAg nanoshells.

Myxovirus protein A (MxA) is a biomarker that can be used to distinguish between viral and bacterial infections. While MxA lateral flow assays (LFAs) have been successfully used for viral vs. bacterial differential diagnosis for children, the clinically relevant level of MxA for adults has been reported to be 100 times lower, which is too low for traditional LFAs. We present results applying the use of surface enhanced Raman spectroscopy (SERS) to detect MxA. AuAg nanoshells (AuAg NSs) were used to enhance the Raman signal of mercaptobenzoic acid (4-MBA), enabling readout by SERS. The AuAg NSs were conjugated to antibodies for the biomarker of interest, resulting in a "nanotag", that could be used in a dipstick immunoassay for detection. We first optimized the nanotag parameters using anti-human IgG/human IgG as a model antibody/antigen system, and then demonstrated detection of MxA using anti-MxA antibodies. We show that SERS readout of immunoassays for MxA can quantify MxA levels at clinically relevant levels for adult viral infection.

Low-Cost Strategy for the Development of a Rapid Electrochemical Assay for Bacteria Detection Based on AuAg Nanoshells

A low-cost strategy for the simple and rapid detection of bacterial cells in biological matrixes is presented herein. Escherichia coli and Salmonella typhimurium were chosen as model bacteria for t...

Tunable electrochemistry of gold-silver alloy nanoshells

The widespread and increasing interest in enhancing biosensing technologies by increasing their sensitivities and lowering their costs has led to the exploration and application of complex nanomaterials as signal transducers and enhancers. In this work, the electrochemical properties of monodispersed AuAg alloy nanoshells (NSs) with finely tunable morphology, composition, and size are studied to assess their potential as electroactive labels. The controlled corrosion of their silver content, caused by the oxidizing character of dissolved oxygen and chlorides of the electrolyte, allows the generation of a reproducible electrochemical signal that is easily measurable through voltammetric techniques. Remarkably, the underpotential deposition of dissolved Ag+ catalyzed on AuAg NS surfaces is observed and its dependence on the nanoparticle morphology, size, and elemental composition is studied, revealing a strong correlation between the relative amounts of the two metals. The highest catalytic activity is found at Au/Ag ratios higher than ≈ 10, showing how the synergy between both metals is necessary to trigger the enhancement of Ag+ reduction. The ability of AuAg NSs to generate an electrocatalytic current without the need for any strong acid makes them an extremely promising material for biosensing applications.

Time- and Size-Resolved Plasmonic Evolution with nm Resolution of Galvanic Replacement Reaction in AuAg Nanoshells Synthesis

The rational design of advanced nanomaterials with enhanced optical properties can be reached only with the profound thermodynamic and kinetic understanding of their synthetic processes. In this work, the synthesis of monodisperse AuAg nanoshells with thin shells and large voids is achieved through the development of a highly reproducible and robust methodology based on the galvanic replacement reaction. This is obtained thanks to the systematic identification of the role played by the different synthetic parameters involved in the process (such as surfactants, co-oxidizers, complexing agents, time, and temperature), providing an unprecedented control over the material’s morphological and optical properties. Thus, the time- and size-resolved evolution of AuAg nanoshells surface plasmon resonance band is described for 15, 30, 60, 80, 100, and 150 nm-sized particles spanning almost through the entire visible spectrum. Its analysis reveals a four-phase mechanism coherent with the material’s morphological tra...

Gold and silver quantification from gold-silver nanoshells in HaCaT cells

A method to determine total gold (Au) and/or silver (Ag) elemental concentrations from gold nanoparticles, Au-Ag nanoshells (NS) and silica coated Au-Ag nanoshells was developed, evaluated and validated. Samples were mineralized in a mixture of concentrated aqua regia and hydrofluoric acid at 65 °C for 4 h. Mineralized solutions were diluted and standard solutions were prepared in aqua regia 5%. ICP-MS analysis was performed with or without the use of a reaction cell (CRC). For the determination of elemental concentrations of nanopowders and test suspensions, the average recovery was 99 ± 2% and 101 ± 2% for gold and silver respectively. The repeatability was evaluated by the Relative Standard Deviation (RSD). The overall analytical RSD was ≤4% (n = 3) and the RSD associated to ICP-MS analysis was ≤2% (n = 10). The limits of detection were 0.005 and 0.002 μg(element) L−1 (analyzed solution), and the limits of quantitation 0.017 and 0.005 μg(element) L−1 (analyzed solution), for 197Au and 109Ag respectively. The Ag/Au mass ratios of the NS in the different samples considered were all equal to (0.93 ± 0.04). From this information, the average thickness of gold and silver layers in the nanoshells was deduced, being 7.5 ± 0.5 and 23 ± 3 nm respectively. Finally, the developed method was successfully applied to in vitro studies to evaluate NS cellular uptake in HaCaT keratinocyte cells confirming the method robustness toward biological medium. Experiments in cell culture medium gave coherent concentrations, 70–100% of uncoated or silica-coated NS being recovered, distributed between the culture medium and the cells (internalized). The analytical repeatability (over the whole procedure, or that of the ICP-MS analysis only) remains in the same order of magnitude as in test suspensions. Minimum concentrations less than or equal to 1 μg(element) g−1(suspension) were determined with the same accuracy.

Near-IR-Absorbing Gold Nanoframes with Enhanced Physiological Stability and Improved Biocompatibility for In Vivo Biomedical Applications.

This paper describes the synthesis of near-infrared (NIR)-absorbing gold nanoframes (GNFs) and a systematic study comparing their physiological stability and biocompatibility with those of hollow Au-Ag nanoshells (GNSs), which have been used widely as photothermal agents in biomedical applications because of their localized surface plasmon resonance (LSPR) in the NIR region. The GNFs were synthesized in three steps: galvanic replacement, Au deposition, and Ag dealloying, using silver nanospheres (SNP) as the starting material. The morphology and optical properties of the GNFs were dependent on the thickness of the Au coating layer and the degree of Ag dealloying. The optimal GNF exhibited a robust spherical skeleton composed of a few thick rims, but preserved the distinctive LSPR absorbance in the NIR region-even when the Ag content within the skeleton was only 10 wt %, 4-fold lower than that of the GNSs. These GNFs displayed an attractive photothermal conversion ability and great photothermal stability, and could efficiently kill 4T1 cancer cells through light-induced heating. Moreover, the GNFs preserved their morphology and optical properties after incubation in biological media (e.g., saline, serum), whereas the GNSs were unstable under the same conditions because of rapid dissolution of the considerable silver content with the shell. Furthermore, the GNFs had good biocompatibility with normal cells (e.g., NIH-3T3 and hepatocytes; cell viability for both cells: >90%), whereas the GNSs exhibited significant dose-dependent cytotoxicity (e.g., cell viability for hepatocytes at 1.14 nM: ca. 11%), accompanied by the induction of reactive oxygen species. Finally, the GNFs displayed good biocompatibility and biosafety in an in vivo mouse model; in contrast, the accumulation of GNSs caused liver injury and inflammation. Our results suggest that GNFs have great potential to serve as stable, biocompatible NIR-light absorbers for in vivo applications, including cancer detection and combination therapy.

AuBr2–-Engaged Galvanic Replacement for Citrate-Capped Au–Ag Alloy Nanostructures and Their Solution-Based Surface-Enhanced Raman Scattering Activity

This work demonstrates that AuBr2– can serve as a suitable AuI precursor in the galvanic replacement reaction with Ag nanoparticles (NPs), especially for preparing citrate-capped Au–Ag alloy nanostructures. A cost-effective recipe for stable AuBr2– solution was presented by the reduction of AuBr4– derived from the ion exchange between AuCl4– and optimal amount of Br–. This recipe successfully suppressed the disproportionation of Au+ without the aid of concentrated salt solution, ensuring the colloidal stability of the citrate-capped Ag NPs during the replacement reaction. A comparative study was then performed on the structural evolution of citrate-capped Ag NPs during the replacement reaction with AuBr2– and AuCl4–. Uniform Au–Ag nanoshells were gradually formed when AuBr2– was employed, while porous Au–Ag nanoframes are generated by using AuCl4–. Moreover, the as-obtained AuBr2– solution can also be extended to the synthesis of complex nanostructures, such as Au@Au–Ag nanorattles. Finally, an improved s...

Self-assembled Monolayer Mediated Surface Environment Modification of Poly(vinylpyrrolidone)-Coated Hollow Au-Ag Nanoshells for Enhanced Loading of Hydrophobic Drug and Efficient Multimodal Therapy.

Hollow Au-Ag bimetallic nanoshell possessing hydrophobic interior space and hydrophilic exterior surface was prepared and its application as a chemo-thermo-gene therapeutic agent based on its high payload of multiple drugs having different water solubility was demonstrated. The multifunctional drug delivery system is based on the hydrophobic interior created by the self-assembled monolayer (SAM) of hexanethiol onto the inner surface of the hollow metallic nanoshells whereas the outer surface was mostly coated by hydrophilic biocompatible polymer. The nanoshells having surface environment modified by hexanethiol SAMs provided high capacity both for hydrophilic DNAzyme (Dz) to induce gene silencing and for hydrophobic SN38 (7-ethyl-10-hydroxycamptothecin), anticancer drug. The release of the loaded Dz and SN38 was independently triggered by an acidic environment and by photothermal temperature elevation upon irradiation, respectively. The chemo-thermo-gene multitherapy based on the present nanoshells having modified surface environment showed high efficacy in quantitative cell-based assays using Huh7 human liver cell containing hepatitis C viral NS3 gene replicon RNA.

The Surprising in Vivo Instability of Near-IR-Absorbing Hollow Au–Ag Nanoshells

Photothermal ablation based on resonant illumination of near-infrared-absorbing noble metal nanoparticles that have accumulated in tumors is a highly promising cancer therapy, currently in multiple clinical trials. A crucial aspect of this therapy is the nanoparticle size for optimal tumor uptake. A class of nanoparticles known as hollow Au (or Au–Ag) nanoshells (HGNS) is appealing because near-IR resonances are achievable in this system with diameters less than 100 nm. However, in this study, we report a surprising finding that in vivo HGNS are unstable, fragmenting with the Au and the remnants of the sacrificial Ag core accumulating differently in various organs. We synthesized 43, 62, and 82 nm diameter HGNS through a galvanic replacement reaction, with nanoparticles of all sizes showing virtually identical NIR resonances at ∼800 nm. A theoretical model indicated that alloying, residual Ag in the nanoparticle core, nanoparticle porosity, and surface defects all contribute to the presence of the plasmon resonance at the observed wavelength, with the major contributing factor being the residual Ag. While PEG functionalization resulted in stable nanoparticles under laser irradiation in solution, an anomalous, strongly element-specific biodistribution observed in tumor-bearing mice suggests that an avid fragmentation of all three sizes of nanoparticles occurred in vivo. Stability studies across a wide range of pH environments and in serum confirmed HGNS fragmentation. These results show that NIR resonant HGNS contain residual Ag, which does not stay contained within the HGNS in vivo. This demonstrates the importance of tracking both materials of a galvanic replacement nanoparticle in biodistribution studies and of performing thorough nanoparticle stability studies prior to any intended in vivo trial application.

Mechanistic study of substrate-based galvanic replacement reactions

The sacrificial templates used in galvanic replacement reactions dictate the properties of the hollow metal nanostructures formed. Here, we demonstrate that substrate-based Au-Ag nanoshells with radically altered properties are obtained by merely coating silver templates with an ultrathin layer of gold prior to their insertion into the reaction vessel. The so-formed nanoshells exhibit much smoother surfaces, a higher degree of crystallinity and are far more robust. Dealloying the nanoshells results in the first demonstration of substrate-based nanocages. Such cages exhibit a well-defined pattern of geometric openings in directions corresponding to the {111}-facets of the starting template material. The ability to engineer the cage geometry through adjustments to the orientational relationship between the crystal structure of the starting template and that of underlying substrate is demonstrated. Together these discoveries provide the framework to advance our understanding of the mechanisms governing substratebased galvanic replacement reactions. Open image in new window

Femtogram detection of cytokines in a direct dot-blot assay using SERS microspectroscopy and hydrophilically stabilized Au–Ag nanoshells

Rapid parallel detection of two cytokines (IL-6 and IL-8) with femtogram sensitivity in a simple direct dot-blot assay is demonstrated. The microspectroscopic SERS acquisition scheme employs rationally designed, hydrophilically stabilized Au-Ag nanoshells as SERS labels, which are optimized for signal enhancement upon red laser excitation.

Substrate-based galvanic replacement reactions carried out on heteroepitaxially formed silver templates

Galvanic replacement reactions have been widely used to transform solution dispersed silver template structures into intricate nanoshell geometries. Here, we report on the use of these same reactions to form hollow substrate-supported Au-Ag nanoshells from silver templates having a heteroepitaxial relationship with the underlying single crystal substrate. The structures obtained exhibit a nanohut geometry, show highly tunable plasmonic properties and are formed as periodic arrays using a lithography-free technique. When removed from the substrate the inverted nanohuts appear as nanobowls with a notch in the rim. The study lays the groundwork for wafer-based devices utilizing nanoshells located at site-specific locations. Open image in new window

Size-dependent transformation from Ag templates to Au–Ag nanoshells via galvanic replacement reaction in organic medium

The transformation from Ag templates to Au–Ag nanoshells via galvanic replacement reaction with HAuCl4 was systematically studied in an organic medium in the presence of oleylamine. Decahedral (~43 nm in size) and triangular prism (~53 nm in edge length) Ag templates transformed into equiaxed and triangular prismatic Au–Ag nanoshells, respectively. The first step involved structural and morphological changes from Ag templates to Au–Ag nanoshells with an interior cavity. In the second step, the growth of the shells continued through the deposition of Au. The shell thickness increased from ~5 to ~10 nm for the equiaxed Au–Ag nanoshells (~39-nm interior cavity) and ~5 to ~8 nm for the triangular prismatic Au–Ag nanoshells (~52-nm interior edge length). Oleylamine not only served as a surfactant but also removed AgCl precipitates and reduced HAuCl4. For the nanoshells derived from the ~20-nm Ag decahedrons, further reaction in excess HAuCl4 collapsed the nanoshells into Au-rich solid fragments. However, the nanoshells derived from the ~43-nm Ag decahedrons, the nanoshell structure not only persisted in excess HAuCl4, but its shell thickness also increased. The size-dependent transformation of these nanoshells is discussed.

Differential SERS Activity of Gold and Silver Nanostructures Enabled by Adsorbed Poly(vinylpyrrolidone)

We report that poly(vinylpyrrolidone) (PVP), a common stabilizer of colloidal dispersions of noble metal nanostructures, has a dramatic effect on their surface-enhanced Raman scattering (SERS) activity and enables highly selective SERS detection of analytes of various type and charge. Nanostructures studied include PVP-stabilized Au-Ag nanoshells synthesized by galvanic exchange reaction of citrate-reduced Ag nanoparticles (NPs), as well as solid citrate-reduced Ag and Au NPs, both before and after stabilization with PVP. All nanostructures were characterized in terms of their size, surface plasmon resonance wavelength, surface charge, and chemical composition. While the SERS activities of the parent citrate-reduced Ag and Au NPs are similar for rhodamine 6G (R6G) and 1,2-bis(4-pyridyl)ethylene (BPE) at various pH values, PVP-stabilized nanostructures demonstrate large differences in SERS enhancement factors (EFs) between these analytes depending on their chemical nature and protonation state. At pH values higher than BPE's pK(a2) of 5.65, where the analyte is largely unprotonated, the PVP-coated Au-Ag nanoshells showed a high SERS EF of >10(8). In contrast, SERS EFs were 10(3)- to 10(5)-fold lower for the protonated form of BPE at lower pH values, or for the usually highly SERS-active cationic R6G. The differential SERS activity of PVP-stabilized nanostructures is a result of discriminatory binding of analytes within-adsorbed PVP monolayer and a subsequent increase of analyte concentration at the nanostructure surface. Our experimental and theoretical quantum chemical calculations show that BPE binding with PVP-stabilized Au-Ag nanoshells is stronger when the analyte is in its unprotonated form as compared to its cationic, protonated form at a lower pH.

Noble metal nanostructure both as a SERS nanotag and an analyte probe

We report nanotags encapsulated in Au-Ag nanostructure that are active in surface-enhanced Raman scattering. The Au-Ag nanoshells are filled with Ag via citrate reduction, entrapping label molecules in the process. The nanotags can be used for quantitative SERS measurements with the label molecules as internal reference.