SERS detection of foodborne pathogens in beverage with Au nanostars.

The aim of this study is to develop a simple but rapid method for the determination of foodborne pathogens in complex matrices (beverages) by surface enhanced Raman spectroscopy (SERS) combined with Au nanostar solid-phase substrates. The star-shaped singlet Au nanostructure was formed on the surface of a stainless steel sheet by chemical replacement reaction. Rhodamine 6G verified the sensitivity and reproducibility of this substrate, and the relative standard deviations of the SERS intensity at 1312 cm-1, 1364 cm-1, and 1510 cm-1 displacements were 3.40%, 5.64%, and 3.48%, respectively. By detecting four pathogens in beverage samples on Au nanostar substrates, the utility of the SERS assay was demonstrated, while the combination of principal component analysis (PCA) and hierarchical cluster analysis (HCA) further enabled the isolation and identification of pathogens. The results of spiked beverages were validated in conventional culture identification and Vitek 2 Compact biochemical identification system experiments. Thus, this research demonstrated that Au nanostar substrates can be effectively utilized for the recognition of pathogenic bacteria and have immense promise to advance the progress of quick detection of foodborne pathogens and food safety.

Embedding Raman Tags between Au Nanostar@Nanoshell for Multiplex Immunosensing.

Novel Raman tags with various reporter molecules embedded in between gold nanostar (AuNS) and gold nanoshell are developed, showing significantly enhanced surface-enhanced Raman scattering intensity compared to gold nanoparticle-based composites. Immunoassay using these AuNS@tag@shell structures is highly specific with sensitivity down to 0.1 pg mL-1 , and is capable of multiplex detection, making them highly promising for biosensing applications.

Development of Au Nanoparticle Two-Dimensional Assemblies Dispersed with Au Nanoparticle-Nanostar Complexes and Surface-Enhanced Raman Scattering Activity.

We recently found that polyvinylpyrrolidone (PVP)-protected metal nanoparticles dispersed in water/butanol mixture spontaneously float to the air/water interface and form two-dimensional assemblies due to classical surface excess theory and Rayleigh-Bénard-Marangoni convection induced by butanol evaporation. In this study, we found that by leveraging this principle, a unique structure is formed where hetero gold nanospheres (AuNPs)/gold nanostars (AuNSs) complexes are dispersed within AuNP two-dimensional assemblies, obtained from a mixture of polyvinylpyrrolidone-protected AuNPs and AuNSs that interact electrostatically with the AuNPs. These structures were believed to form as a result of AuNPs/AuNSs complexes formed in the water/butanol mixture floating to the air/water interface and being incorporated into the growth of AuNP two-dimensional assemblies. These structures were obtained by optimizing the amount of mixed AuNSs, with excessive addition resulting in the formation of random three-dimensional network structures. The AuNP assemblies dispersed with AuNPs/AuNSs complexes exhibited significantly higher Raman (surface-enhanced resonance Raman scattering: SERRS) activity compared to simple AuNP assemblies, while the three-dimensional network structure did not show significant SERRS activity enhancement. These results demonstrate the excellent SERRS activity of AuNP two-dimensional assemblies dispersed with hetero AuNPs/AuNSs complexes.

Ultrasensitive dual-enhanced sandwich strategy for simultaneous detection of Escherichia coli and Staphylococcus aureus based on optimized aptamers-functionalized magnetic capture probes and graphene oxide-Au nanostars SERS tags.

In this work, a novel surface-enhanced Raman scattering (SERS) sandwich strategy biosensing platform has been established for simultaneously detecting Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). Fe3O4@SiO2-Au nanocomposites (NCs) with varying amounts of Au nanocrystals were prepared, and the effect of interparticle gaps on SERS activity was studied by finite-difference time-domain (FDTD) method. The optimal magnetic SERS-active substrates (FS-A5) were functionalized with the specific aptamers to act as capture probes. Meanwhile, graphene oxide-Au nanostars (GO-Au NSs) decorated with Raman reporters and aptamers were used as SERS tags. The loading density of Au NSs on GO was tuned to change the number of SERS active sites. In this proposal, E. coli and S. aureus were first captured by capture probes and then bound with SERS tags to form a sandwich-like structure, which caused enhanced electromagnetic field because of the dual enhancement strategy. Under optimal conditions, SERS platform could detect E. coli and S. aureus simultaneously, and the detection limit was as low as 10 cfu/mL. Our sandwich assay-based dual-enhanced SERS platform provides a new idea for simultaneously detecting multiple pathogens with high selectivity and sensitivity, and thus will have more hopeful prospects in the field of food safety.

Multi-dimensional plasmonic coupling system for efficient enrichment and ultrasensitive label-free SERS detection of bilirubin based on graphene oxide-Au nanostars and Au@Ag nanoparticles.

Rapid and sensitive detection of free bilirubin (BR) is essential for early diagnosis of jaundice and other hepatobiliary diseases. Inspired by sandwich immunoassay strategy, a multi-dimensional plasmonic coupling SERS platform composed of graphene oxide-Au nanostars nanocomposites (GANS NCs) and Au@Ag nanoparticles (NPs) was designed for label-free detection of BR. Specifically, GANS NCs were first prepared, and their excellent SERS activity was ascribed to synergistic enhancement effect of electromagnetic enhancement and chemical enhancement. Furthermore, SERS spectroscopy was used to monitor the adsorption process of BR. Subsequently, secondary reinforcing Au@Ag NPs were directly added, ultimately resulting in a multi-dimensional plasmonic coupling effect. The SERS enhancing mechanism of coupled system was discussed through electromagnetic field simulations. Interestingly, the high-density hotspots generated by strong plasmonic coupling in GANS-Au@Ag substrate could lead to more extraordinary SERS enhancing behavior compared to GANS NCs. Sensing efficiency of the SERS platform was examined by BR with a detection limit down to 10-11 M. Besides, GANS-Au@Ag NCs performed high uniformity and reproducibility. This work not only opens up a new avenue for construction of multi-dimensional plasmonic coupling system, but also offers a new biosensing technology for label-free diagnosis of BR-related diseases, thereby expecting to be applied in clinical diagnosis.

Multifunctional Plasmon-Tunable Au Nanostars and Their Applications in Highly Efficient Photothermal Inactivation and Ultra-Sensitive SERS Detection.

The development and application in different fields of multifunctional plasmonic nanoparticles (NPs) have always been research hotspots. Herein, multi-tip Au nanostars (NSs) with an anisotropic structure were fabricated for the photothermal therapy (PTT) of bacteria and surface-enhanced Raman scattering (SERS) detection of pollutants. The size and localized surface plasmon resonance (LSPR) characteristics of Au NSs were adjusted by varying Au seed additions. In addition, photothermal conversion performance of Au NSs with various Au seed additions was evaluated. Photothermal conversion efficiency of Au NSs with optimal Au seed additions (50 µL) was as high as 28.75% under 808 nm laser irradiation, and the heat generated was sufficient to kill Staphylococcus aureus (S. aureus). Importantly, Au NSs also exhibited excellent SERS activity for the 4-mercaptobenzoic acid (4-MBA) probe molecule, and the local electromagnetic field distribution of Au NSs was explored through finite-difference time-domain (FDTD) simulation. As verified by experiments, Au NSs' SERS substrate could achieve a highly sensitive detection of a low concentration of potentially toxic pollutants such as methylene blue (MB) and bilirubin (BR). This work demonstrates a promising multifunctional nanoplatform with great potential for efficient photothermal inactivation and ultra-sensitive SERS detection.

In Situ Direct Monitoring of the Morphological Transformation of Single Au Nanostars Induced by Iodide through Dual-Laser Dark-Field Microscopy: Unexpected Mechanism and Sensing Applications.

Single nanoparticle imaging is a significant technique to help reveal the reaction mechanism and provides insight into the nanoparticle transformation. Here, we monitor the in situ morphological transformation of Au nanostars (GNSs) induced by iodide (I-) in real time using dark-field microscopy (DFM) with 638 nm red (R) and 534 nm green (G) laser coillumination. The two lasers are selected because the longitudinal localized surface plasmon resonance of GNSs is located at 638 nm and that for GNSs after transformation is at 534 nm. Interestingly, I- can interact with GNSs directly without the engagement of other reagents, and upon increasing I- concentrations, GNSs undergo color changes from red to orange, yellow, and green under DFM. Accordingly, green/red channel intensities (G/R ratios) are extracted by obtaining red and green channel intensities of single nanoparticles to weigh the morphological changes and quantify I-. A single nanoparticle sensor is constructed for I- detection with a detection limit of 6.9 nM. Finally, a novel mechanism is proposed to elucidate this shape transformation. I- absorbed onto the surface of GNSs binds with Au atoms to form AuI-, lowering the energy of its bond with other Au atoms, which facilitates the diffusion of this atom across the nanoparticle surface to low-energy sites at the concaves, thus deforming to spherical Au nanoparticles.

Serum-based surface-enhanced Raman spectroscopy combined with PCA-RCKNCN for rapid and accurate identification of lung cancer.

Early and precise diagnosis of lung cancer is critical for a better prognosis. However, it is still a challenge to develop an effective strategy for early precisely diagnose and effective treatments. Here, we designed a label-free and highly accurate classification serum analytical platform for identifying mice with lung cancer. Specifically, the microarray chip integrated with Au nanostars (AuNSs) array was employed to measure the surface-enhanced Raman scattering (SERS) spectra of serum of tumor-bearing mice at different stages, and then a recognition model of SERS spectra was constructed using the principal component analysis (PCA)-representation coefficient-based k-nearest centroid neighbor (RCKNCN) algorithm. The microarray chip can realize rapid, sensitive, and high-throughput detection of SERS spectra of serum. RCKNCN based on the PCA-generated features successfully differentiated the SERS spectra of serum of tumor-bearing mice at different stages with a classification accuracy of 100%. The most prominent spectral features for distinguishing different stages were captured in PCs loading plots. This work not only provides a practical SERS chip for the application of SERS technology in cancer screening, but also provides a new idea for analyzing the feature of serum at the spectral level.

Live Cell Poration by Au Nanostars to Probe Intracellular Molecular Composition with SERS.

A new type of flat substrate has been used to visualize structures inside living cells by surface-enhanced Raman scattering (SERS) and to study biochemical processes within cells. The SERS substrate is formed by stabilized aggregates of gold nanostars on a glass microscope slide coated with a layer of poly (4-vinyl pyridine) polymer. This type of SERS substrate provides good cell adhesion and viability. Au nanostars' long tips can penetrate the cell membrane, allowing it to receive the SERS signal from biomolecules inside a living cell. The proposed nanostructured surfaces were tested to study, label-free, the distribution of various biomolecules in cell compartments.

Magnetite-Supported Gold Nanostars for the Uptake and SERS Detection of Tetracycline.

Magnetite nanoparticles (MNPs) decorated with gold nanostars (AuNSs) have been prepared by using a seed growth method without the addition of surfactants or colloidal stabilizers. The hybrid nanomaterials were investigated as adsorbents for the uptake of tetracycline (TC) from aqueous solutions and subsequent detection using surface-enhanced Raman scattering (SERS). Several parameters were investigated in order to optimize the performance of these hybrid platforms on the uptake and SERS detection of TC, including variable pH values and the effect of contact time on the removal of TC. The spatial distribution of TC and AuNS on the hybrid composites was accomplished by coupling SERS analysis with Raman imaging studies, allowing also for the determination of the detection limit for TC when dissolved in ultrapure water (10 nM) and in more complex aqueous matrices (1 µM). Attempts were also made to investigate the adsorption modes of the TC molecules at the surface of the metal NPs by taking into account the enhancement of the Raman bands in these different matrices.

Gold nanostars: Benzyldimethylammonium chloride-assisted synthesis, plasmon tuning, SERS and catalytic activity.

Fabrication of Au nanostars (AuNSs) can expand the application range of Au nanoparticles because of their high electron density and localized surface plasmon resonance (LSPR) on branches. Exploiting this potential requires further refinement of length of the branches and radius of their tips. To this end, we successfully synthesized AuNSs with uniform and sharply-pointed branches by combining benzyldimethylammonium chloride (BDAC) and cetyltrimethylammonium bromide (CTAB) at low BDAC/CTAB ratios. Once mixed with CTAB, BDAC lowers the critical micelle concentration (CMC) for quick formation of the micelles, which provides favorable growth templates for AuNSs formation. Besides, BDAC increases the concentration of Cl(-), which favors Ag(+) in adsorbing on Au facets. This feature is crucial for the yield boosting and synergic shape control of AuNSs regardless of types of Au seeds used. Use of less amounts of seeds as the center of nucleation benefited sharper and longer growth of the branches. AuNSs exhibited excellent enhancement of surface-enhanced Raman scattering (SERS) intensities as the result of high electron density localized at the tips; however, the enhancement degree varied in accordance with the size of branches. In addition, AuNSs showed high catalytic performance toward the reduction of 4-nitrophenol (4-NP) to 4-aminophenol (4-AP). Efficient catalysis over AuNSs originates from their corners, stepped surfaces and high electron density at the tips.

Intracellular surface-enhanced Raman scattering probes based on TAT peptide-conjugated Au nanostars for distinguishing the differentiation of lung resident mesenchymal stem cells.

Lung resident mesenchymal stem cells (LR-MSCs) are important regulators of pathophysiological processes including tissue repair and fibrosis, inflammation, angiogenesis and tumor formation. Therefore, increasing attention has focused on the functional differentiation of LR-MSCs. However, the distinction between the undifferentiated and differentiated LR-MSCs, which are closely related and morphologically similar, is difficult to achieve by conventional methods. In this study, by employing the TAT Peptide-conjugated Au nanostars (AuNSs) as an intracellular probe, we developed a method for the identification of LR-MSC differentiation by surface-enhanced Raman scattering (SERS) spectroscopy. SERS spectra were analyzed using principal component analysis (PCA) that allowed unambiguous distinction of subtypes and monitoring of component changes during cellular differentiation. Furthermore, to ascertain whether co-culture with alveolar epithelial type II (ATII) cells and incubation with transform growth factor (TGF)-ß were involved in regulating the differentiation of LR-MSCs, we investigated the protein expression levels of epithelial markers and fibroblastic markers on LR-MSCs. Our results demonstrated that co-culture with ATII cells or incubation with TGF-ß could induce the differentiation of LR-MSCs as confirmed by SERS analysis, a method that is capable of noninvasive characterization of and distinction between subtypes of LR-MSCs during differentiation. We have provided a new tool that may facilitate stem cell research.

Hyaluronic acid-modified Fe3O4@Au core/shell nanostars for multimodal imaging and photothermal therapy of tumors.

Development of multifunctional theranostic nanoplatforms for diagnosis and therapy of cancer still remains a great challenge. In this work, we report the use of hyaluronic acid-modified Fe3O4@Au core/shell nanostars (Fe3O4@Au-HA NSs) for tri-mode magnetic resonance (MR), computed tomography (CT), and thermal imaging and photothermal therapy of tumors. In our approach, hydrothermally synthesized Fe3O4@Ag nanoparticles (NPs) were used as seeds to form Fe3O4@Au NSs in the growth solution. Further sequential modification of polyethyleneimine (PEI) and HA affords the NSs with excellent colloidal stability, good biocompatibility, and targeting specificity to CD44 receptor-overexpressing cancer cells. With the Fe3O4 core NPs and the star-shaped Au shell, the formed Fe3O4@Au-HA NSs are able to be used as a nanoprobe for efficient MR and CT imaging of cancer cells in vitro and the xenografted tumor model in vivo. Likewise, the NIR absorption property enables the developed Fe3O4@Au-HA NSs to be used as a nanoprobe for thermal imaging of tumors in vivo and photothermal ablation of cancer cells in vitro and xenografted tumor model in vivo. This study demonstrates a unique multifunctional theranostic nanoplatform for multi-mode imaging and photothermal therapy of tumors, which may find applications in theranostics of different types of cancer.

Effect of Au NanoStars/collagen matrix material on oxidative stress damage in ventricular myocytes of newborn rats / 军事医学

Objective To verify antioxidation of Au NanoStars/collagen ( AuNSs/Col ) for ventricular myocytes of newborn rats(NRVMs) by in vitro studies.Methods (1)Different concentrations of AuNSs/Col composite materials were created.The optimum concentration of the material was selected by Live /dead staining and Cell Counting Kit-8 (CCK-8) and Col was used for subsequent experiments .( 2 ) NRVMs were randomly divided into Col group , AuNSs/Col group, H2O2-induced Col group, and H2O2-induced AuNSs/Col group.After 6 h treatment, apoptotic cell morphology and early cell apoptosis rate were observed with Annexin Ⅴ-FITC/propidium iodide ( PI)/4′,6-diamidino-2-phenylindole ( DAPI) and the expressions of apoptosis related proteins-B-cell lymphoma-2 ( Bcl-2 ) and Bcl-2 associated x protein ( Bax ) were detected by Western blotting .Results ( 1 ) Both Live/dead and CCK-8 experiments indicated that the AuNSs/Col composite material with 0.1 mg/ml was nontoxicity to NRVMs and could further promote their proliferation .(2) Compared with the uninduced group , the early apoptosis rate of the Col group and the AuNSs /Col group after H2O2 induction was significantly increased , while the Bcl-2/Bax ratio was decreased , indicating that the oxidative stress damage model was established.After H2O2 induction, compared with the Col group , the early apoptosis rate of the AuNSs/Col group was decreased , but the Bcl-2/Bax ratio was increased .Conclusion AuNSs/Col composite material has protective effect on the oxidative damage of cardiomyocytes cultured in vitro.