Graphene-based high-efficiency surface-enhanced Raman scattering-active platform for sensitive and multiplex DNA detection

We have developed a surface-enhanced Raman scattering (SERS)-active substrate based on gold nanoparticle-decorated chemical vapor deposition (CVD)-growth graphene and used it for multiplexing detection of DNA. Due to the combination of gold nanoparticles and graphene, the Raman signals of dye were dramatically enhanced by this novel substrate. With the gold nanoparticles, DNA capture probes could be easily assembled on the surface of graphene films which have a drawback to directly immobilize DNA. This platform exhibits extraordinarily high sensitivity and excellent specificity for DNA detection. A detection limit as low as 10 pM is obtained. Importantly, two different DNA targets could be detected simultaneously on the same substrate just using one light source.     

Interaction of gold nanoparticles with Pfu DNA polymerase and effect on polymerase chain reaction

The interaction of gold nanoparticles with Pfu DNA polymerase has been investigated by a number of biological, optical and electronic spectroscopic techniques. Polymerase chain reaction was performed to show gold nanoparticles' biological effect. Ultraviolet-visible and circular dichroism spectra analysis were applied to character the structure of Pfu DNA polymerase after conjugation with gold nanoparticles. X-ray photoelectron spectroscopy was used to investigate the bond properties of the polymerase-gold nanoparticles complex. The authors demonstrate that gold nanoparticles do not affect the amplification efficiency of polymerase chain reaction using Pfu DNA polymerase, and Pfu DNA polymerase displays no significant changes of the secondary structure upon interaction with gold nanoparticles. The adsorption of Pfu DNA polymerase to gold nanoparticles is mainly through Au-NH(2) bond and electrostatic interaction. These findings may have important implications regarding the safety issue as gold nanoparticles are widely used in biomedical applications.     

One-pot polymerase chain reaction with gold nanoparticles for rapid and ultrasensitive DNA detection

We report a novel method for rapid, colorimetric detection of a specific deoxyribonucleic acid (DNA) sequence by carrying out a polymerase chain reaction in the presence of gold nanoparticles functionalized with two primers. Extension of the primers when the target DNA is present as a template during the polymerase chain reaction process affords the complementary sequences on the gold nanoparticle surfaces and results in the formation of gold nanoparticle aggregates with a concomitant color change from red to pinkish/purple. This method provides a convenient and straightforward solution for ultrasensitive DNA detection without any further post-treatment of the polymerase chain reaction products being necessary, and is a promising tool for rapid disease diagnostics and gene sequencing.     

Impedance sensing of DNA binding drugs using gold substrates modified with gold nanoparticles

Interfacial interactions between immobilized DNA probes and DNA-specific sequence binding drugs were investigated using impedance spectroscopy toward the development of a novel biosensing scheme. The impedance measurements are based on the charge-transfer kinetics of the [Fe(CN)6]3-/4- redox couple. Compared to bare gold surfaces, the immobilization of DNA and then the DNA-drug interaction on electrode surfaces altered the capacitance and the interfacial electron resistance and thus diminished the charge-transfer kinetics by reducing the active area of the electrode or by preventing the redox species from approaching the electrode. Electrochemical deposition of gold nanoparticles on a gold electrode surface showed significant improvement in sensitivity. DNA-capped gold nanoparticles on electrodes act as selective sensing interfaces with tunable sensitivity due to higher amounts of DNA probes and the concentric orientation of the DNA self-assembled monolayer. The specificity of the interactions of two classical minor groove binders, mythramycin, a G-C specific-DNA binding anticancer drug, netropsin, an A-T specific-DNA binding drug and an intercalator, nogalamycin on AT-rich DNA-modified substrate and GC-rich DNA-modified substrate are compared. Using gold nanoparticle-deposited substrates, impedance spectroscopy resulted in a 20-40-fold increase in the detection limit. Arrays of deposited gold nanoparticles on gold electrodes offered a convenient tool to subtly control probe immobilization to ensure suitably adsorbed DNA orientation and accessibility of other binding molecules.     

Optically controlled thermal management on the nanometer length scale

The manipulation of polymers and biological molecules or the control of chemical reactions on a nanometer scale by means of laser pulses shows great promise for applications in modern nanotechnology, biotechnology, molecular medicine or chemistry. A controllable, parallel, highly efficient and very local heat conversion of the incident laser light into metal nanoparticles without ablation or fragmentation provides the means for a tool like a 'nanoreactor', a 'nanowelder', a 'nanocrystallizer' or a 'nanodesorber'. In this paper we explain theoretically and show experimentally the interaction of laser radiation with gold nanoparticles on a polymethylmethacrylate (PMMA) layer (one-photon excitation) by means of different laser pulse lengths, wavelengths and pulse repetition rates. To the best of our knowledge this is the first report showing the possibility of highly local (in a 40?nm range) regulated heat insertion into the nanoparticle and its surroundings without ablation of the gold nanoparticles. In an earlier paper we showed that near-infrared femtosecond irradiation can cut labeled DNA sequences in metaphase chromosomes below the diffraction-limited spot size. Now, we use gold as well as silver-enhanced gold nanoparticles on DNA (also within chromosomes) as energy coupling objects for femtosecond laser irradiation with single-and two-photon excitation. We show the results of highly localized destruction effects on DNA that occur only nearby the nanoparticles.     

The In Vivo Radiosensitizing Effect of Gold Nanoparticles Based MRI Contrast Agents

Owing to the high atomic number (Z) of gold element, the gold nanoparticles appear as very promising radiosensitizing agents. This character can be exploited for improving the selectivity of radiotherapy. However, such an improvement is possible only if irradiation is performed when the gold content is high in the tumor and low in the surrounding healthy tissue. As a result, the beneficial action of irradiation (the eradication of the tumor) should occur while the deleterious side effects of radiotherapy should be limited by sparing the healthy tissue. The location of the radiosensitizers is therefore required to initiate the radiotherapy. Designing gold nanoparticles for monitoring their distribution by magnetic resonance imaging (MRI) is an asset due to the high resolution of MRI which permits the accurate location of particles and therefore the determination of the optimal time for the irradiation. We recently demonstrated that ultrasmall gold nanoparticles coated by gadolinium chelates (Au@DTDTPA‐Gd) can be followed up by MRI after intravenous injection. Herein, Au@DTDTPA and Au@DTDTPA‐Gd were prepared in order to evaluate their potential for radiosensitization. Comet assays and in vivo experiments suggest that these particles appear well suited for improving the selectivity of the radiotherapy. The dose which is used for inducing similar levels of DNA alteration is divided by two when cells are incubated with the gold nanoparticles prior to the irradiation. Moreover, the increase in the lifespan of tumor bearing rats is more important when the irradiation is performed after the injection of the gold nanoparticles. In the case of treatment of rats with a brain tumor (9L gliosarcoma, a radio‐resistant tumor in a radiosensitive organ), the delay between the intravenous injection and the irradiation was determined by MRI.     

Hybrid composite material based on polythiophene derivative nanofibers modified with gold nanoparticles for optoelectronics applications

Conjugated polymers have been extensively applied as active materials in nanostructured platforms for optical and electrical devices. The incorporation of metal nanoparticles (NPs) into the polymer-based platform arises as a strategy to develop novel hybrid functional nanocomposites with enhanced electrical and optical properties. However, efficient and simple processing routes to produce such nanocomposites are still on demand. In this work, we present an effective route to obtain functional nanocomposites based on electrospun nanofibers coated with gold nanoparticles, displaying interesting optical and electrical properties. Polymethyl methacrylate (PMMA) electrospun nanofibers doped with poly(3-hexyl thiophene-2,5-diyl) (P3HT) were obtained by the electrospinning technique, and displayed a strong red emission centered at 650 nm assigned to P3HT. Such nanofibers were deposited on to fluorine-doped tin oxide electrodes and with modified with gold nanoparticles (AuNPs) in order to produce hybrid composite materials. The performance of electrodes modified with PMMA/P3HT-AuNPs composite material was evaluated by impedance spectroscopy and revealed an enhancement of electron transfer kinetics, which indicates it as a potential platform for optical and electrochemical (bio)sensors.     

Inhibition of HSV‐1 Attachment,Entry, and Cell‐to‐Cell Spread by Functionalized Multivalent Gold Nanoparticles

The use of modified nanoparticles in interactions with biological targets is attracting rapidly increasing attention. In this Full Paper, the application of gold nanoparticles capped with mercaptoethanesulfonate (Au‐MES NPs) as effective inhibitors of Herpes simplex virus type 1 infection based on their ability to mimic cell‐surface‐receptor heparan sulfate is described. Mechanistic studies reveal that Au‐MES NPs interfere with viral attachment, entry, and cell‐to‐cell spread, thereby preventing subsequent viral infection in a multimodal manner. The ligand multiplicity achieved with carrier nanoparticles is crucial in generating polyvalent interactions with the virus at high specificity, strength, and efficiency. Such multivalent‐nanoparticle‐mediated inhibition is a promising approach for alternative antiviral therapy.     

Interfacial dispersion of poly(N-isopropylacrylamide)/ gold nanocomposites

The nanoparticle dispersity and interfacial property could be considered as a basis of their further application in the nanostructured materials. In this paper, the dispersity and interfacial phenomena of poly(N-isopropylacrylamide) modified gold nanoparticles were investigated. Firstly, such polymer/gold nanocomposites were demonstrated to have a good dispersity in water, tetrahydrofuran, alcohols and also chloroform, so they were used to entrap fluorescent dye-labelled lipids in chloroform as nanocontainers and subsequently delivery the fluorescent lipids into water as nanocarriers. Secondly, when the nanocomposites in water/chloroform mixture were heated above 35 degrees C, the nanocomposite particles could be partially transferred from water into chloroform across the interface, and they would come back into water again as cooling, displaying a reversible thermal response. Moreover, such polymer/gold nanocomposites at the immiscible water/toluene fluids preferred to assembly into 2-dimensional membranes with variable density at the water/oil interface. The special dispersion properties of the poly(N-isopropylacrylamide)/gold nanocomposites provide many potentials in the future.     

Ionogel-templated synthesis and organization of anisotropic gold nanoparticles

Photochemical reduction of tetrachloroaurate (AuCl4-) ions in the highly constrained aqueous domains of a nanostructured ionogel template, formed via self-assembly of the ionic liquid 1-decyl-3-methylimidazolium chloride (C10mim+Cl-) in water, results in the formation of anisotropic gold nanoparticles with a variety of sizes and morphologies, which include previously unattainable trigonal prismatic nanorods. Unexpectedly, small-angle X-ray scattering studies of the Au-ionogel composite reveal that the in situ formation of the nanoparticles increases the mesoscopic order of the ionogel, which results in its conversion to a near-monodomain structure. The findings demonstrate that nanostructured, ionic liquid-based gels can be used to template the formation of new nanoparticle morphologies with technologically important optical, electronic, and catalytic properties. It may also be possible to design soft templates that permit the fabrication of highly ordered nanoparticle array-hydrogel composites, thereby enabling control and tuning of the collective properties of the encapsulated nanoparticles.     

Characterization of different sequences of DNA on si substrate by atomic force microscopy and gold nanoparticle labeling

In this paper, different sequences of single-strand DNA modified on Si substrate were studied taking advantages of the high resolution of atomic force microscopy (AFM) and signal enhancement of gold nanoparticles. Two sequences of single-strand DNA, as a model, were immobilized on Si substrate and hybridized with their sequence-complementary DNA molecules modified respectively with two sizes of gold nanoparticles. The surface of Si substrate was characterized through detecting the size and coverage of gold nanoparticles by AFM. Results demonstrated that different sizes of gold nanoparticles represented different sequences of DNA immobilized on the substrate. Density and distribution of DNA on Si substrate can be investigated by AFM imaging using gold nanoparticles as topographic markers. Compared to other sensitive methods such as fluorescence energy transfer, X-ray photoelectron, and radiolabeling experiments, this approach is advantageous in terms of high spatial resolution in sub-micrometer scale. This new method will be beneficial in the characterization of DNA immobilized on chip surfaces.     

Single-step co-deposition of nanostructured tungsten oxide supported gold nanoparticles using a gold–phosphine cluster complex as the gold precursor

The use of a molecular gold organometallic cluster in chemical vapour deposition is reported, and it is utilized, together with a tungsten oxide precursor, for the single-step co-deposition of (nanostructured) tungsten oxide supported gold nanoparticles (NPs). The deposited gold-NP and tungsten oxide supported gold-NP are highly active catalysts for benzyl alcohol oxidation; both show higher activity than SiO₂ supported gold-NP synthesized via a solution-phase method, and tungsten oxide supported gold-NP show excellent selectivity for conversion to benzaldehyde.     

Layer-by-layer assembled charge-trap memory devices with adjustable electronic properties

We describe a versatile approach for preparing flash memory devices composed of polyelectrolyte/gold nanoparticle multilayer films. Anionic gold nanoparticles were used as the charge storage elements, and poly(allylamine)/poly(styrenesulfonate) multilayers deposited onto hafnium oxide (HfO2)-coated silicon substrates formed the insulating layers. The top contact was formed by depositing HfO2 and platinum. In this study, we investigated the effect of increasing the number of polyelectrolyte and gold nanoparticle layers on memory performance, including the size of the memory window (the critical voltage difference between the 'programmed' and 'erased' states of the devices) and programming speed. We observed a maximum memory window of about 1.8 V, with a stored electron density of 4.2 x 1012 cm-2 in the gold nanoparticle layers, when the devices consist of three polyelectrolyte/gold nanoparticle layers. The reported approach offers new opportunities to prepare nanostructured polyelectrolyte/gold nanoparticle-based memory devices with tailored performance.     

Gold nanoparticles with asymmetric polymerase chain reaction for colorimetric detection of DNA sequence

We developed a novel strategy for rapid colorimetric analysis of a specific DNA sequence by combining gold nanoparticles (AuNPs) with an asymmetric polymerase chain reaction (As-PCR). In the presence of the correct DNA template, the bound oligonucleotides on the surface of AuNPs selectively hybridized to form complementary sequences of single-stranded DNA (ssDNA) target generated from As-PCR. DNA hybridization resulted in self-assembly and aggregation of AuNPs, and a concomitant color change from ruby red to blue-purple occurred. This approach is simpler than previous methods, as it requires a simple mixture of the asymmetric PCR product with gold colloid conjugates. Thus, it is a convenient colorimetric method for specific nucleic acid sequence analysis with high specificity and sensitivity. Most importantly, the marked color change occurs at a picogram detection level after standing for several minutes at room temperature. Linear amplification minimizes the potential risk of PCR product cross-contamination. The efficiency to detect Bacillus anthracis in clinical samples clearly indicates the practical applicability of this approach.     

Synthesis of 16-Mercaptohexadecanoic acid capped gold nanoparticles and their immobilization on a substrate

Controlled assembly of nanoparticles on substrates is a promising path to develop miniaturized electronic and optical devices. Among the important issues to be addressed in this area include immobilization of the nanoparticles on substrates in order to ensure that the system is robust. In this work, 16-mercaptohexadecanoic acid (16-MHDA) capped gold nanoparticles with a narrow size distribution have been synthesized through a single phase synthesis method and subsequently immobilized on to silicon surface through covalent molecular assembly. Fourier transform infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS) confirmed the absence of unreacted thiol in the synthesized gold nanoparticles. Presence of gold nanoparticles on Si surface after the immobilization process was confirmed through XPS. Cross-sectional high resolution transmission electron microscopy (HR-TEM) images provide direct evidence that the particles are indeed anchored to the silicon surface. The formation of uniform-sized and separated acid functionalized gold nanoparticles and their immobilization on to Si provide a basis for further nano-structuring.     

Nanostructure characterization of polymer-stabilized gold nanoparticles and nanofilms derived from green synthesis

The fabrication and characterization of gold (Au) nanostructured materials draws significant attention because of their distinctive properties and their technological applications. The first objective of this study is to fabricate polymer-stabilized Au nanoparticles and nanofilms (PAN) through a cost effective and green synthetic methodology. In this study, the gold trication (Au³⁺) can be spontaneously converted into metallic gold atom using a non-toxic reductant (ascorbic acid). The ultrafine Au clusters were formed and stabilized through metallic bonds in the colloidal suspension, which was then deposited on a micro-glass or polymer-bead substrate to prepare thin films. It was found that ascorbic acid was the best reducing agent due to its rapid rate, spontaneity of reaction, and its non-toxic nature. In order to prevent aggregation of the nanoparticles, a dispersing agent (gum Arabic) was used. The second objective of this study was to analyze the PAN using a number of state-of-the-art instrumentation techniques and analytical approaches, such as X-ray powder diffraction (XRD), atomic force microscopy (AFM), scanning and transmission electron microscopy (SEM and TEM), ultraviolet–visible (UV–Vis) spectroscopy, and ZetaPALS. These techniques were applied to evaluate specific properties of the PAN, such as characterization of its crystalline phase, surface topology, characteristic plasmon, particle size distribution, and stability. From this study, it can be concluded that the ultrafine Au nanoparticles and uniform films were obtained using the green chemistry method. The ultrafine Au particles are highly stabilized and monodispersed as demonstrated by their high absolute value of zeta potential.     

Aluminum nanostructured films as substrates for enhanced fluorescence in the ultraviolet-blue spectral region

Particulate aluminum films of varied thicknesses were deposited on quartz substrates by thermal evaporation. These nanostructured substrates were characterized by scanning electron microscopy (SEM). With the increase of aluminum thickness, the films progress from articulate toward smooth surfaces as observed by SEM images. To date, metal-enhanced fluorescence (MEF) has primarily been observed in the visible - NIR wavelength region using silver or gold island films or roughened surfaces. We now show that fluorescence could also be enhanced in the ultraviolet-blue region of the spectrum using nanostructured aluminum films. Two probes, one in the ultraviolet and another one in the blue spectral region, have been chosen for the present study. We observed increased emission, decrease in fluorescence lifetime, and increase in photostability of a DNA base analogue 2-aminopurine and a coumarin derivative (7-HC) in 10-nm spin-casted poly(vinyl alcohol) film on Al nanostructured surfaces. The fluorescence enhancement factor depends on the thickness of the Al films as the size of the nanostructures formed varies with Al thickness. Both probes showed increased photostability near aluminum nanostructured substrates, which is consistent with the shorter lifetime. Our preliminary studies indicate that Al nanostructured substrates can potentially find widespread use in MEF applications particularly in the UV-blue spectral regime. Furthermore, these Al nanostructured substrates are very stable in buffers that contain chloride salts compared to usual silver colloid-based substrates for MEF, thus furthering the usefulness of these Al-based substrates in many biological assays where high concentration of salts are required. Finite-Difference Time-Domain calculations were also employed to study the enhanced near-fields induced around aluminum nanoparticles by a radiating fluorophore, and the effect of such enhanced fields on the fluorescence enhancement observed was discussed.     

Nano‐biosensor based on reduced graphene oxide and gold nanoparticles,for detection of phenylketonuria‐associated DNA mutation

Phenylketonuria (PKU)‐associated DNA mutation in newborn children can be harmful to his health and early detection is the best way to inhibit consequences. A novel electrochemical nano‐biosensor was developed for PKU detection, based on signal amplification using nanomaterials, e.g. gold nanoparticles (AuNPs) decorated on the reduced graphene oxide sheet on the screen‐printed carbon electrode. The fabrication steps were checked by field emission scanning electron microscope imaging as well as cyclic voltammetry analysis. The specific alkanethiol single‐stranded DNA probes were attached by self‐assembly methodology on the AuNPs surface and Oracet blue was used as an intercalating electrochemical label. The results showed the detection limit of 21.3 fM and the dynamic range of 80–1200 fM. Moreover, the selectivity results represented a great specificity of the nano‐biosensor for its specific target DNA oligo versus other non‐specific sequences. The real sample simulation was performed successfully with almost no difference than a synthetic buffer solution environment.Inspec keywords: biosensors, nanosensors, nanoparticles, graphene compounds, gold, nanomedicine, DNA, molecular biophysics, biomedical equipment, electrochemical sensors, electrochemical electrodes, field emission scanning electron microscopy, voltammetry (chemical analysis), self‐assembly, biochemistryOther keywords: reduced graphene oxide, gold nanoparticles, phenylketonuria‐associated DNA mutation, newborn children, electrochemical nanobiosensor, signal amplification, nanomaterials, reduced graphene oxide sheet, screen‐printed carbon electrode, field emission scanning electron microscopy imaging, cyclic voltammetry, alkanethiol single‐stranded DNA probes, self‐assembly methodology, Oracet blue, intercalating electrochemical label, Au‐CO     

A DNA duplex with extremely enhanced thermal stability based on controlled immobilization on gold nanoparticles

The effect of DNA loadings on the thermal stability of DNA duplex immobilized on gold nanoparticles has been investigated. The modestly loaded duplexes on the gold nanoparticles showed enhanced thermal stability, as compared to that of the free duplex (without gold nanoparticles). However, the highly loaded duplex showed stability similar to that of free duplex. The stability could be controlled over a wide temperature range simply by varying the salt concentration (over 50 degrees C). Additionally, the gold nanoparticles with modestly loaded oligonucleotides could be used as nanoprobes for effective and fast strand exchange reactions, based on the increased thermal stability of the immobilized duplex. These results indicate that the interaction between the duplex and the nanoparticle surface plays an important role in determining the stability of the duplex.