Femtomolar electrochemical detection of DNA hybridization using hollow polyelectrolyte shells bearing silver nanoparticles

The preparation, and use as electrochemical labels, of polyelectrolyte shells bearing Ag nanoparticles is described. Their potential for highly sensitive detection is demonstrated. The shells are prepared by layer-by-layer self-assembly around templates (500 nm diameter) which are then dissolved. The shells can be opened and closed by adjustment of solution pH, and this process is utilized to encapsulate Ag nanoparticles, chiefly by adsorption to the inner walls of the capsules. Based on absorbance, TEM and voltammetric measurements, the highest loading achieved is approximately 78 Ag particles per capsule. The Ag capsules are used via biotin-avidin binding as labels for the detection of DNA hybridization, following acid dissolution and then detection of the Ag (+) by ASV. A 30-mer sequence specific to Escherichia coli is measured at DNA-modified screen-printed electrodes with a detection limit of approximately 25 fM, which corresponds to the detection of 4.6 fg ( approximately 3 x 10 (5) molecules) in the 20 microL analyte sample. A 200 fM target containing a single mismatch gives a significantly (<74%) lower response than 200 fM of complementary target; 60 pM of noncomplementary target gives a negligible response.     

Fluorescence enhancement of modified silver nanoparticles

Surface enhanced fluorescence (SEF) effect of acridine orange fluorophore in the proximity of silver nanoparticles (NPs) has been investigated experimentally in the aqueous solution system. It was found that the SEF effect could be influenced by the distribution of the NPs and the separation between the fluorophore molecule and metal surface. The fluorescence enhancement was improved significantly when Ag NPs was capped with 4-Aminothiophenol (PATP) that was acted as an isolating layer between the metal surface and fluorophore molecules. The results suggest that a proper distribution of metallic NPs and proper separation between fluorophore molecule and the particle surface are important for obtaining an optimal SEF effect.     

Single DNA molecule detection using nanopipettes and nanoparticles

Single DNA molecules labeled with nanoparticles can be detected by blockades of ionic current as they are translocated through a nanopipette tip formed by a pulled glass capillary. The nanopipette detection technique can provide not only tools for detection and identification of single DNA and protein molecules but also deeper insight and understanding of stochastic interactions of various biomolecules with their environment.     

Gram-scale synthesis and biofunctionalization of silica-coated silver nanoparticles for fast colorimetric DNA detection

A direct silica-coating method has been developed for the gram-scale synthesis of well-dispersed Ag@SiO(2) nanoparticles. Subsequent surface functionalization via the well-established silica surface chemistry provided arching points for straightforward bioconjugation with amino-terminated oligonucleotides. Fast hybridization kinetics of the resulting robust oligo-modified Ag@SiO(2) nanoprobes with complementary target oligonucleotides render themselves very useful for the fast colorimetric DNA detection based on the sequence-specific hybridization properties of DNA. Additionally, the reliable protocols developed in this study for preparing and functionalizing Ag@SiO(2) nanoparticles can be readily extended to other silica-coated nanoparticles, which can also provide a specific platform for the covalent attachment of biomolecules such as amino-rich proteins, enzymes, or amino-terminated oligonucleotides for diverse bioapplications.     

One-step synthesis of silver nanoparticles at the air-water interface using different methods

Silver nanoparticles were synthesized in a one-step process at the air-AgNO(3) aqueous solution interface under Langmuir monolayers of 5,10,15,20-tetra-4-oxy(2-stearic acid) phenyl porphyrin (TSPP) at room temperature by using different methods including UV-light irradiation, ambient light irradiation, and formaldehyde gas reduction. It was found that parallel aligned one-dimensional (1D) chains composed of discrete silver nanoparticles with the size of 3-5?nm were formed under UV-light irradiation for a short time, while large areas of uniform silver spherical nanoparticles were formed under natural daylight illumination for several days or by formaldehyde gas treatment for several hours. The average size of the spherical nanoparticles ranges from 6.88 ± 0.46 to 11.10 ± 1.47?nm, depending on the experimental conditions. The 1D chains formed under UV-light irradiation result from the templating effect of parallel aligned linear supramolecular arrays formed by TSPP at the air-water interface, and rapid nucleation and growth of the nanoparticles. The formation of the uniform silver nanoparticles under daylight illumination or by formaldehyde gas treatment, however, should be ascribed to a kinetically controlled growth process of the nanoparticles.     

Fluorescence enhancement using silver nanotriangle arrays

We describe the fabrication of silver nanotriangle array using angle resolved nanosphere lithography and utilizing the same for enhancing fluorescence. The well established nanosphere lithography is modified by changing the angle of deposition between the nanosphere mask and the beam of silver being deposited resulting in nanotriangles of varying surface area and density. The 470 nm plasmon resonance wavelength of the substrate was determined using minimum reflectivity method which closely matches with excitation wavelength of the fluorophore. Ten times enhancement in fluorescence emission intensity is obtained from fluorescein isothiocyanate coated on top of silver nanotriangle array separated by a spacer layer of poly vinyl alcohol as compared to glass. The enhanced fluorescence emission is attributed to the increase in local field enhancement.     

Green synthesis of silver nanoparticles using tannins

Colloidal silver nanoparticles were prepared by rapid green synthesis using different tannin sources as reducing agent viz. chestnut (CN), mangrove (MG) and quebracho (QB). The aqueous silver ions when exposed to CN, MG and QB tannins were reduced which resulted in formation of silver nanoparticles. The resultant silver nanoparticles were characterized using UV-Visible, X-ray diffraction (XRD), scanning electron microscopy (SEM/EDX), and transmission electron microscopy (TEM) techniques. Furthermore, the possible mechanism of nanoparticles synthesis was also derived using FT-IR analysis. Spectroscopy analysis revealed that the synthesized nanoparticles were within 30 to 75 nm in size, while XRD results showed that nanoparticles formed were crystalline with face centered cubic geometry.     

Enzyme colorimetric assay using unmodified silver nanoparticles

Colorimetric assay based on the unique surface plasmon resonance properties of metallic nanoparticles has received considerable attention in bioassay due to its simplicity, high sensitivity, and low cost. Most of colorimetric methods previously reported employed gold nanoparticles (GNPs) as sensing elements. In this work, we develop a sensitive, selective, simple, and label-free colorimetric assay using unmodified silver nanoparticle (AgNP) probes to detect enzymatic reactions. Enzymatic reactions concerning adenosine triphosphate (ATP) dephosphorylation by calf intestine alkaline phosphatase (CIAP) and peptide phosphorylation by protein kinase A (PKA) were studied. In the absence of the enzymes, unreacted ATP could protect AgNPs from salt-induced aggregation, whereas in the presence of the enzymes, the reaction product of ATP (i.e., adenosine for CIAP and ADP for PKA) could not. Via our method, dephosphorylation and phosphorylation could be readily detected by the color change of AgNPs, with a detection limit of 1 unit/mL for CIAP and a detection limit of 0.022 unit/mL for PKA. More importantly, the enzymatic inhibition by inhibitors and enzymatic activity in complex biological fluids could also be realized. This work is an important step toward a colorimetric assay using AgNPs and might provide a promise for enzyme assay in realistically complex systems and for screening of different enzyme inhibitors in future.     

Biosynthesis of silver nanoparticles using sun-dried mulberry leaf

The biosynthesis of silver nanoparticles (AgNPs) has been successfully conducted by reduction of silver nitrate with sun-dried mulberry leaf. Such AgNPs have been characterized by UV-visible spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, high-resolution transmission electron microscopy (HRTEM) and atomic force microscopy (AFM). The results showed that such dispersed, uniform and spherical AgNPs would not aggregate under high-concentration NaCl solution and have good antibacterial activity. It was suggested that the polyol components (such as polyhydroxylated alkaloids) and protein residues of mulberry leaf should be mainly responsible for the stabilization of AgNPs. Such AgNPs produced by the environmentally friendly method have the potential for use in antibacterial and medical applications.     

Functional finishing of cotton fabrics using silver nanoparticles

We have reported a novel in situ synthesis protocol for silver nanoparticles onto cotton fabrics. Here, cotton fabric immersed in silver nitrate solution is autoclaved at 15 psi, 121 degrees C for 15 min. At this temperature and pressure, the aldehyde terminal of starch (residual size material on cotton fabric) reduced the silver nitrate to silver metal and simultaneously stabilized the nanoparticles on fabric itself. The UV-visible absorption spectrum of both cotton fabrics and bath solution showed a typical absorption peak at 420 nm corresponding to the surface plasmon resonance of silver nanoparticles. With the help of transmission electron micrographs, the average size of the dislodged silver nanoparticles in water is calculated to be 20.9 +/- 13.7 nm. This silver nanoparticles impregnated cotton fabrics showed excellent antibacterial activity against Staphylococcus aureus and bacteriostasis activity against Klebsiella pneumoniae. Also, silver nanoparticles impregnated fabrics expressed significant UV-protection capability in comparison with the untreated fabrics.     

Fluorescence enhancement of silver nanoparticle hybrid probes and ultrasensitive detection of IgE

An ultrasensitive protein assay method was developed based on silver nanoparticle (AgNP) hybrid probes and metal-enhanced fluorescence. Two aptamer based silver nanoparticles, Aptamer/Oligomer-A/Cy3-modified AgNPs (Tag-A) and Aptamer/Oligomer-B/Cy3-modified AgNPs (Tag-B) were hybridized to form a silver nanoparticle aggregate that produced a red shift and broadening of the Localized Surface Plasmon Resonance (LSPR) peak. The enhanced fluorescence resulted from the increased content of Cy3 molecules and their emission resonance coupled to the broadened localized surface plasmon (LSP) of AgNP aggregate. The separation distance between Cy3 and AgNPs was 8 nm which was the most optimal for metal enhanced fluorescence and the separation distance between adjacent AgNPs was about 16 nm and this was controlled by the lengths of oligomer-A and oligomer-B. The protein array was prepared by covalently immobilizing capture antibodies on aldehyde-coated slide. After addition of protein IgE sample, two kinds of aptamer-modified AgNPs (Tag-A and Tag-B) were employed to specifically recognize IgE and form the AgNP aggregate on the arrays based on their hybridization. The detection property of the aptamer-modified AgNP aggregate was compared to two other modified aptamer-based probes, aptamer-modified Cy3 and Tag-A. The modified AgNP hybrid probe (Tag-A and Tag-B) showed remarkable superiority in both sensitivity and detection limit due to the formed AgNP aggregate. The new hybrid probe also produced a wider linear range from 0.49 to 1000 ng/mL with the detection limit reduced to 40 pg/mL (211 fM). The presented method showed that the newly designed strategy of combining aptamer-based nanomaterials to form aggregates results in a highly sensitive optical detection method based on localized surface plasmon.     

Ultrasensitive flow injection chemiluminescence detection of DNA hybridization using signal DNA probe modified with Au and CuS nanoparticles

A novel and sensitive flow injection chemiluminescence assay for sequence-specific DNA detection based on signal amplification with nanoparticles (NPs) is reported in the present work. The "sandwich-type" DNA biosensor was fabricated with the thiol-functionalized capture DNA first immobilized on an Au electrode and hybridized with one end of target DNA, the other end of which was recognized with a signal DNA probe labeled with CuS NPs and Au NPs on the 3'- and 5'-terminus, respectively. The hybridization events were monitored by the CL intensity of luminol-H2O2-Cu(2+) after the cupric ions were dissolved from the hybrids. We demonstrated that the incorporation of Au NPs in this sensor design significantly enhanced the sensitivity and the selectivity because a single Au NP can be loaded with hundreds of signal DNA probe strands, which were modified with CuS NPs. The ratios of Au NPs, signal DNA probes, and CuS NPs modified on the gold electrode were approximately 1/101/103. A preconcentration process of cupric ions performed by anodic stripping voltammetry technology further increased the sensor performance. As a result of these two combined effects, this DNA sensor could detect as low as femtomolar target DNA and exhibited excellent selectivity against two-base mismatched DNA. Under the optimum conditions, the CL intensity was increased with the increase of the concentration of target DNA in the range of 2.0 x 10(-14)-2.0 x 10(-12) M. A detection limit of 4.8 x 10(-15) M target DNA was achieved.     

Biofabrication of silver nanoparticles using bacteria from mangrove swamp

The last decade has observed a rapid advancement in utilising biological system towards bioremediation of metal ions in the form of respective metal nanostructures or microstructures. The process may also be adopted for respective metal nanoparticle biofabrication. Among different biological methods, bacteria‐mediated method is gaining great attention for nanoparticle fabrication due to their eco‐friendly and cost‐effective process. In the present study, silver nanoparticle (AgNP) was synthesised via continuous biofabrication using Aeromonas veronii, isolated from swamp wetland of Sunderban, West Bengal, India. The biofabricated AgNP was further purified to remove non‐conjugated biomolecules using size exclusion chromatography, and the purified AgNPs were characterised using UV–visible spectroscopy, X‐ray diffraction, field emission scanning electron microscopy and transmission electron microscopy (TEM). Additionally, the presence of proteins as capping and stabilising agents was confirmed by the amide‐I and amide‐II peaks in the spectra obtained using attenuated total reflection Fourier transform infrared spectroscopy. The size of biofabricated AgNP was 10–20 nm, as observed using TEM. Additionally, biofabricated AgNP shows significant antibacterial potential against E. coli and S. aureus. Hence, biofabricated AgNP using Aeromonas veronii, which found resistant to a significant concentration of Ag ion, showed enhanced antimicrobial activity compared to commercially available AgNP.Inspec keywords: silver, nanoparticles, microorganisms, nanofabrication, purification, chromatography, ultraviolet spectra, visible spectra, X‐ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, proteins, molecular biophysics, biochemistry, molecular configurations, attenuated total reflection, Fourier transform infrared spectra, particle size, antibacterial activity, biomedical materials, nanomedicineOther keywords: capping agents, stabilising agents, amide‐I peaks, amide‐II peaks, attenuated total reflection Fourier transform infrared spectroscopy, antibacterial potential, E. coli, S. aureus, Aeromonas veronii, antimicrobial activity, size 10 nm to 20 nm, Ag, proteins, TEM, transmission electron microscopy, field emission scanning electron microscopy, X‐ray diffraction, UV‐visible spectroscopy, size exclusion chromatography, nonconjugated biomolecules, purification, swamp wetland, Aeromonas veronii, cost‐effective process, eco‐friendly, bacteria‐mediated method, biological methods, metal nanoparticle biofabrication, microstructures, metal nanostructures, metal ions, bioremediation, biological system, mangrove swamp, bacteria, silver nanoparticles     

Synthesis of silver nanoparticles by chemical and biological methods and their antimicrobial properties

Biogenic synthesis of nanoparticles offers an attractive alternate to chemical synthesis methods. Various hazard free, eco-friendly methods of synthesis of silver nanoparticles are in operation. In chemical reduction methods, the reducing agent is a chemical solution, whereas in biological ones, the collection of enzymes, especially nitrate reductase, plays this role. The highest antibacterial activity of silver nanoparticles synthesised by chemical and biological methods was found in Staphylococcus aureus and Escherichia coli. The paper aims to discuss some fundamental issues about non-biological methods and benefits about biological methods for silver nanoparticles synthesis and their antibacterial studies.     

Quantitative enhanced Raman scattering of labeled DNA from gold and silver nanoparticles

Surface-enhanced resonance Raman scattering (SERRS) from silver nanoparticles using 514.5-nm excitation has been shown to offer huge potential for applications in highly sensitive multiplexed DNA assays. If the technique is to be applied to real biological samples and integrated with other methods, then the use of gold nanoparticles and longer wavelengths of excitation are desirable. The data presented here demonstrate that dye-labeled oligonucleotide sequences can be directly detected by SERRS using gold nanoparticles in a quantitative manner for the first time. The performance of gold and silver nanoparticles as SERRS substrates was assessed using 514.5-, 632.8-, and 785-nm excitation and a range of 13 commercially available dye-labeled oligonucleotides. The quantitative response allowed the limit of detection to be determined for each case and demonstrates that the technique is highly effective, sensitive, and versatile. The possibility of excitation at multiple wavelengths further enhances the multiplexing potential of the technique. The importance of effectively combining the optical properties of the nanoparticle and the dye label is demonstrated. For example, at 632.8-nm excitation, the dye BODIPY TR-X and gold nanoparticles make a strong SERRS combination with very little background fluorescence. This study allows the choice of nanoparticle and dye label for particular experimental setups, and significantly expands the applicability of enhanced Raman scattering for use in many disciplines.     

Highly sensitive SERS detection of As3+ ions in aqueous media using glutathione functionalized silver nanoparticles

A highly sensitive surface-enhanced Raman scattering (SERS) platform for the selective trace analysis of As(3+) ions was reported based on glutathione (GSH)/4-mercaptopyridine (4-MPY)-modified silver nanoparticles (AgNPs). Here, GSH conjugated on the surface of AgNPs for specifical binding with As(3+) ions in aqueous solution through As-O linkage and 4-MPY was used as a Raman reporter. When As(3+) ions were added to the system, the binding of As(3+) with GSH resulted in the aggregation of AgNPs, and excellent Raman signal of 4-MPY reporters was obtained which can reflect the concentration of As(3+) indirectly. Under optimal assay conditions, the limit of detection (LOD) was estimated to be as low as 0.76 ppb, which is lower than the WHO defined limit (10 ppb), and an excellent linear range of 4-300 ppb was obtained. The practical application had been carried out for determination of As(3+) in real water samples.     

Broadband enhancement in thin-film amorphous silicon solar cells enabled by nucleated silver nanoparticles

Recently plasmonic effects have gained tremendous interest in solar cell research because they are deemed to be able to dramatically boost the efficiency of thin-film solar cells. However, despite of the intensive efforts, the desired broadband enhancement, which is critical for real device performance improvement, has yet been achieved with simple fabrication and integration methods appreciated by the solar industry. We propose in this paper a novel idea of using nucleated silver nanoparticles to effectively scatter light in a broadband wavelength range to realize pronounced absorption enhancement in the silicon absorbing layer. Since it does not require critical patterning, experimentally these tailored nanoparticles were achieved by the simple, low-cost and upscalable wet chemical synthesis method and integrated before the back contact layer of the amorphous silicon thin-film solar cells. The solar cells incorporated with 200 nm nucleated silver nanoparticles at 10% coverage density clearly demonstrate a broadband absorption enhancement and significant superior performance including a 14.3% enhancement in the short-circuit photocurrent density and a 23% enhancement in the energy conversion efficiency, compared with the randomly textured reference cells without nanoparticles. Among the measured plasmonic solar cells the highest efficiency achieved was 8.1%. The significant enhancement is mainly attributed to the broadband light scattering arising from the integration of the tailored nucleated silver nanoparticles.     

Green synthesis of silver nanoparticles using biopolymers,carboxymethylated-curdlan and fucoidan

There is a growing need in developing a reliable and eco-friendly methodology for the synthesis of metallic nanoparticles, which may be applied for many nanotechnological applications. Natural compounds such as biopolymers are one of the resources which could be used for this purpose. The present study involves the development of a simple, ecological and user-friendly method in synthesizing silver nanoparticles by using carboxymethylated-curdlan or fucoidan as reducing and stabilizing agents. Reduction of silver ions by these biopolymers occurred when heating at 100 °C, led to the formation of silver nanoparticles in the range of 40–80 nm in dimensions. The silver nanoparticles were formed readily within 10–15 min. Morphological observation and characterization of the silver nanoparticles were performed by using dynamic light scattering (DLS), high-resolution transmission electron microscopy (HRTEM), and UV–vis absorption spectrophotometer. The size of silver nanoparticles can be controlled by using different concentrations of carboxymethylated-curdlan, fucoidan or silver nitrate. This way of silver nanoparticles preparation is easy, fast, user-friendly and suitable for large-scale production.     

DNA sequence detection using selective fluorescence quenching of tagged oligonucleotide probes by gold nanoparticles

Simple, fast, economical, and sensitive detection of specific DNA sequences is crucial to pathogen detection and biomedical research. We have designed a novel fluorescent assay for DNA hybridization based on the electrostatic properties of DNA. We exploit the ability to create conditions where single-stranded DNA adsorbs on negatively charged gold nanoparticles while double-stranded DNA does not. Dye-tagged probe sequences have their fluorescence efficiently quenched when they are mixed with gold nanoparticles unless they hybridize with components of the analyte. Subfemtomole amounts of untagged target are detected in minutes using commercially available materials. Target sequences in complex mixtures of DNA and single-base mismatches in DNA sequences are easily detected.