Counting single native biomolecules and intact viruses with color-coded nanoparticles

Nanometer-sized particles such as semiconductor quantum dots and energy-transfer nanoparticles have novel optical properties such as tunable light emission, signal brightness, and multicolor excitation that are not available from traditional organic dyes and fluorescent proteins. Here we report the use of color-coded nanoparticles and dual-color fluorescence coincidence for real-time detection of single native biomolecules and viruses in a microfluidic channel. Using green and red nanoparticles to simultaneously recognize two binding sites on a single target, we demonstrate that individual molecules of genes, proteins, and intact viruses can be detected and identified in complex mixtures without target amplification or probe/target separation. Real-time coincidence analysis of single-photon events allows rapid detection of bound targets and efficient discrimination of excess unbound probes. Quantitative studies indicate that the counting results are remarkably precise when the total numbers of counted molecules are more than 10. The use of bioconjugated nanoparticle probes for single-molecule detection is expected to have important applications in ultrasensitive molecular diagnostics, bioterrorism agent detection, and real-time imaging and tracking of single-molecule processes inside living cells.     

Detecting single viruses and nanoparticles using whispering gallery microlasers

There is a strong demand for portable systems that can detect and characterize individual pathogens and other nanoscale objects without the use of labels, for applications in human health, homeland security, environmental monitoring and diagnostics. However, most nanoscale objects of interest have low polarizabilities due to their small size and low refractive index contrast with the surrounding medium. This leads to weak light-matter interactions, and thus makes the label-free detection of single nanoparticles very difficult. Micro- and nano-photonic devices have emerged as highly sensitive platforms for such applications, because the combination of high quality factor Q and small mode volume V leads to significantly enhanced light-matter interactions. For example, whispering gallery mode microresonators have been used to detect and characterize single influenza virions and polystyrene nanoparticles with a radius of 30 nm (ref. 12) by measuring in the transmission spectrum either the resonance shift or mode splitting induced by the nanoscale objects. Increasing Q leads to a narrower resonance linewidth, which makes it possible to resolve smaller changes in the transmission spectrum, and thus leads to improved performance. Here, we report a whispering gallery mode microlaser-based real-time and label-free detection method that can detect individual 15-nm-radius polystyrene nanoparticles, 10-nm gold nanoparticles and influenza A virions in air, and 30 nm polystyrene nanoparticles in water. Our approach relies on measuring changes in the beat note that is produced when an ultra-narrow emission line from a whispering gallery mode microlaser is split into two modes by a nanoscale object, and these two modes then interfere. The ultimate detection limit is set by the laser linewidth, which can be made much narrower than the resonance linewidth of any passive resonator. This means that microlaser sensors have the potential to detect objects that are too small to be detected by passive resonator sensors.     

Infrared spectroscopic mapping of single nanoparticles and viruses at nanoscale resolution

We demonstrate that scattering near-field microscopy (s-SNOM) can determine infrared "fingerprint" spectra of individual poly(methyl methacrylate) nanobeads and viruses as small as 18 nm. Amplitude and phase spectra are found surprisingly strong, even at a probed volume of only 10(-20) l, and robust in regard to particle size and substrate. This makes infrared spectroscopic s-SNOM a versatile tool for chemical and-in the case of protein-secondary-structure identification.     

High-throughput detection and sizing of individual low-index nanoparticles and viruses for pathogen identification

Rapid, chip-scale, and cost-effective single particle detection of biological agents is of great importance to human health and national security. We report real-time, high-throughput detection and sizing of individual, low-index polystyrene nanoparticles and H1N1 virus. Our widefield, common path interferometer detects nanoparticles and viruses over a very large sensing area, orders of magnitude larger than competing techniques. We demonstrate nanoparticle detection and sizing down to 70 nm in diameter. We clearly size discriminate nanoparticles with diameters of 70, 100, 150, and 200 nm. We also demonstrate detection and size characterization of hundreds of individual H1N1 viruses in a single experiment.     

Near-static dielectric polarization of individual carbon nanotubes

Low-frequency dielectric responses of carbon nanotubes are important for their manipulation, separation, and electronic applications. Here we report the first experimental measurement of near-dc polarization of individual carbon nanotubes by using modified scanning force microscopy techniques. The transverse polarizability of carbon nanotubes is equivalent to solid cylindrical media with a dielectric constant of about 10, irrespective of tube diameter and chirality. The longitudinal polarization is also observed and used to distinguish metallic from semiconducting nanotubes.     

Label-free single-nucleotide polymorphism detection using a cationic tetrahedralfluorene and silica nanoparticles

We developed a simple method that is able to provide label-free sequence-specific DNA detection with single-nucleotide polymorphism (SNP) detection selectivity. This method makes use of both DNA probe immobilized silica nanoparticles and optically amplifying light harvesting molecules. The recognition is accomplished by sequence-specific hybridization between the DNA probes on the silica nanoparticles and the targets of interest. After subsequent treatment with ethidium bromide (EB), a cationic tetrahedralfluorene was added to electrostatically associate with the DNA molecules on the nanoparticle surface, leading to sensitized EB emission via fluorescence resonance energy transfer (FRET). Because of the selective response of the tetrahedralfluorene to intercalated EB, the perfectly matched DNA targets were distinctively differentiated from those with mutations. The presence of tetrahedralfluorene provides improved detection sensitivity and selectivity, as compared to the use of EB alone as a signal reporter. The demonstrated highly selective label-free detection method laid the ground work for the future development of disposable and real-time testing kits in SNP screenings.     

Conduction and dielectric polarization in Se thin films

Se films were prepared by thermal evaporation technique in thickness range 150-8500 Å. X-ray diffraction measurements showed that Se films are in the amorphous state. The ac conductivity and dielectric properties of the amorphous Se films have been investigated in the frequency range 100-100 KHz and 100-400 K temperature range. The ac conductivity σ_ac(ω) is found to be proportional to ω^s where s<1. The temperature dependence of both ac conductivity and the parameter s is reasonably well interpreted by the correlated barrier (CBH) model. The dc conductivity at the room temperature was also studied in the same thickness range. It was concluded that the same mechanism of carrier motion might be dominant in both ac polarization and dc conduction. This carrier transport mechanism might be electronic.     

Angle-suppressed scattering and optical forces on submicrometer dielectric particles

We show that submicrometer silicon spheres, whose polarizabilities are completely given by their two first Mie coefficients, are an excellent laboratory to test effects of both angle-suppressed and resonant differential scattering cross sections. Specifically, outstanding scattering angular distributions, with zero forward- or backward-scattered intensity, (i.e., the so-called Kerker conditions), previously discussed for hypothetical magnetodielectric particles, are now observed for those Si objects in the near infrared. Interesting new consequences for the corresponding optical forces are derived from the interplay, both in and out of resonance, between the electric- and magnetic-induced dipoles.     

Nanometer positioning, parallel alignment, and placement of single anisotropic nanoparticles using hydrodynamic forces in cylindrical droplets

Droplets of liquid drying on a surface with pinned contact area develop an internal hydrodynamic flow that carries entrained particles to the air-liquid-substrate interface. We use this phenomenon in cylindrical, micrometer-sized droplets of large aspect ratio (more than 1000:1) to align, position, and place individual anisotropic nanostructures such as single-walled carbon nanotubes (SWNT). More than 84% of SWNT are aligned in parallel within +/-5 degrees relative to the target axis of alignment. A potential flow model accurately describes and quantifies the statistical variation in the positioning of the nanostructures. We demonstrate for the first time the top-down parallel alignment and placement of individual (unbundled) nanotubes from solution electrically contacted across gold electrodes.     

Label-free detection of single protein molecules and protein-protein interactions using synthetic nanopores

Nanofabricated pores in 20 nm-thick silicon nitride membranes were used to probe various protein analytes as well as to perform an antigen-antibody binding assay. A two-compartment electrochemical cell was separated by a single nanopore, 28 nm in diameter. Adding proteins to one compartment caused current perturbations in the ion current flowing through the pore. These perturbations correlated with both the charge and the size of the protein or of a protein-protein complex. The potential of this nanotechnology for studying protein-protein interactions is highlighted with the sensitive detection of beta-human chorionic gonadotropin, a hormone and clinical biomarker of pregnancy, by monitoring in real time and at a molecular level the formation of a complex between hormones and antibodies in solution. In this form, the assay compared advantageously to immunoassays, with the important difference that labels, immobilization, or amplification steps were no longer needed. In conclusion, we present proof-of-principle that properties of proteins and their interactions can be investigated in solution using synthetic nanopores and that these interactions can be exploited to measure protein concentrations accurately.     

Detection of nanoparticles using optical gradient forces

Abstract

We present a detection scheme for nanoscale particles based on the gradient force and torque near a tightly focused laser beam. The focus affects the path of nanoparticles passing by and a quadrant detector records the particle trajectory. A feedback system continuously adjusts the laser power and thereby prevents the particles from being trapped. Particle size and shape can be assessed by evaluating the time-trace of the quadrant detector signal.     

Label-free plasmonic detection of biomolecular binding by a single gold nanorod

We report the use of individual gold nanorods as plasmonic transducers to detect the binding of streptavidin to individual biotin-conjugated nanorods in real time on a surface. Label-free detection at the single-nanorod level was performed by tracking the wavelength shift of the nanorod-localized surface plasmon resonant scattering spectrum using a dark-field microspectroscopy system. The lowest streptavidin concentration that was experimentally measured was 1 nM, which is a factor of 1000-fold lower than the previously reported detection limit for streptavidin binding by biotinylated single plasmonic nanostructures. We believe that the current optical setup is able to reliably measure wavelength shifts as small as 0.3 nm. Binding of streptavidin at 1 nM concentration induces a mean resonant wavelength shift of 0.59 nm suggesting that we are currently operating at close to the limit of detection of the system.     

Energy transfer in a dielectric medium with allowance for the relaxation polarization

Energy relations of the macroscopic electrodynamics of a dielectric medium are considered with allowance for the relaxation polarization. An expression for the dielectric loss power flux density is obtained in the case of an arbitrary time-dependent electric field. An energy characteristic of the efficiency of insulators for capacitive energy storages is proposed.     

Copper nanoparticles within amorphous and crystalline dielectric matrices

Two approaches are considered for synthesis of Cu nanoparticles within amorphous and crystalline matrices, which allow fabrication of nanoparticles-in-dielectric nanocomposites. The first one is based on the sol–gel technique producing dielectric silica films with copper nanoparticles. The particles provide an optical response of the composite material due to the plasmon resonance band. In the second approach the nanoparticles are produced within the crystalline zeolite matrices, which are able to stabilize both the few-atomic clusters and the nanoparticles. This optically active crystalline material is now introduced into a sol–gel matrix to produce transparent films.     

On the ferroelectric dc nonlinear dielectric permittivity and its relation with the macroscopic polarization

Journal of Materials Science - The relation between the nonlinear dielectric permittivity $$\varepsilon '(E)$$ and the macroscopic polarization P(E) is obtained for ferroelectrics considering a...     

Conduction and dielectric polarization in thin anodic aluminium oxide films

The a.c. polarization of anodic Al₂O₃ films was studied in the thickness range 30–1500 Å and the frequency range 300 Hz to 15 kHz. The interfacial polarization mechanism involving distinct regions separated by flaws and/or voids in the bulk was thought to become dominant with decreasing thickness because of an increasing concentration of defects. The d.c. conductivity at the room temperature was also studied in the same thickness range. The observed linear log J versus E^1/2 characteristics for several thicknesses showed that d.c. conduction in these films obeys the Poole-Frenkel law over a certain range of applied electric fields. Anomalies observed in the electric field and the thickness dependences of the d.c. conductivity are considered to arise from the thickness dependence of the defect concentration. It was concluded that the same mechanism of carrier motion might be dominant in both a.c. polarization and d.c. conduction.     

Electromechanical instability and snap-through instability of dielectric elastomers undergoing polarization saturation

By applying a voltage, electric charge will be induced on the surface of dielectric elastomers. Generally, the charge increases with the level of voltage. When the voltage reaches to a certain value, the charge would not increase any more due to the polarization saturation of dielectric materials. In this paper, a thermodynamic constitutive model, combined both the nonlinear dielectric and hyperelastic behavior as dielectric elastomers undergoing polarization saturation, has been developed. Analytical solutions have been obtained for situations incorporating strain-stiffening effect, electromechanical instability and snap-through instability. The numerical results reveal the marked influence of the extension and polarization saturation limits of elastomer material on its electromechanical instability and the snap-through instability. The developed constitutive model would be helpful in future research of dielectric elastomer based high-performance transducers.     

Electro-optic switching and dielectric spectroscopy studies of ferroelectric liquid crystals with low and high spontaneous polarization

Electro-optic and dielectric studies have been carried out on two ferroelectric liquid crystalline materials: namely, the commercial mixture possessing low spontaneous polarization and pure material with high spontaneous polarization on cells with planar alignment. The calculated values of spontaneous polarization, response time and rotational viscosity were found ~ 240 nC/cm², 20 nC/cm², 1600 μs, 500 μs, 230 kg/ms and 19 kg/ms for high and low polarization ferroelectric liquid crystalline materials, respectively. For the ferroelectric liquid crystal with high polarization, the effect of temperature, frequency and bias field on the dielectric properties have been studied and the obtained results were compared to those obtained on the ferroelectric liquid crystalline mixture with low spontaneous polarization.