The Origin of the Intracellular Silver in Bacteria: A Comprehensive Study using Targeting Gold-Silver Alloy Nanoparticles.

The bactericidal effects of silver nanoparticles (Ag NPs) against infectious strains of multiresistant bacteria is a well-studied phenomenon, highly relevant for many researchers and clinicians battling bacterial infections. However, little is known about the uptake of the Ag NPs into the bacteria, the related uptake mechanisms, and how they are connected to antimicrobial activity. Even less information is available on AgAu alloy NPs uptake. In this work, the interactions between colloidal silver-gold alloy nanoparticles (AgAu NPs) and Staphylococcus aureus (S. aureus) using advanced electron microscopy methods are studied. The localization of the nanoparticles is monitored on the membrane and inside the bacterial cells and the elemental compositions of intra- and extracellular nanoparticle species. The findings reveal the formation of pure silver nanoparticles with diameters smaller than 10 nm inside the bacteria, even though those particles are not present in the original colloid. This finding is explained by a local RElease PEnetration Reduction (REPER) mechanism of silver cations emitted from the AgAu nanoparticles, emphasized by the localization of the AgAu nanoparticles on the bacterial membrane by aptamer targeting ligands. These findings can deepen the understanding of the antimicrobial effect of nanosilver, where the microbes are defusing the attacking silver ions via their reduction, and aid in the development of suitable therapeutic approaches.

Nanoscale Three-Dimensional Charge Density and Electric Field Mapping by Electron Holographic Tomography.

The operation of nanoscale electronic devices is related intimately to the three-dimensional (3D) charge density distributions within them. Here, we demonstrate the quantitative 3D mapping of the charge density and long-range electric field associated with an electrically biased carbon fiber nanotip with a spatial resolution of approximately 5 nm using electron holographic tomography in the transmission electron microscope combined with model-based iterative reconstruction. The approach presented here can be applied to a wide range of other nanoscale materials and devices.

Electron-beam-induced charging of an Al2O3 nanotip studied using off-axis electron holography.

Electrostatic charging of specimens during electron, photon or ion irradiation is a complicated and poorly understood phenomenon, which can affect the acquisition and interpretation of experimental data and alter the functional properties of the constituent materials. It is usually linked to secondary electron emission, but also depends on the geometry and electrical properties of the specimen. Here, we use off-axis electron holography in the transmission electron microscope to study electron-beam-induced charging of an insulating Al2O3 nanotip on a conducting support. The measurements are performed under parallel electron illumination conditions as a function of specimen temperature, electron dose, primary electron energy and surface cleanliness. We observe a lack of reproducibility of charge density measurements after cycling the specimen temperature. Surprisingly, we find both positively and negatively charged regions in closely adjacent parts of the specimen.

Operando transmission electron microscopy of battery cycling: thickness dependent breaking of TiO2 coating on Si/SiO2 nanoparticles.

Conformal coating of silicon (Si) anode particles is a common strategy for improving their mechanical integrity, to mitigate battery capacity fading due to particle volume expansion, which can result in particle crumbling due to lithiation induced strain and excessive solid-electrolyte interface formation. Here, we use operando transmission electron microscopy in an open cell to show that TiO2 coatings on Si/SiO2 particles undergo thickness dependent rupture on battery cycling where thicker coatings crumble more readily than thinner (∼5 nm) coatings, which corroborates the difference in their capacities.

Continuous illumination picosecond imaging using a delay line detector in a transmission electron microscope.

Progress towards analysing transitions between steady states demands improvements in time-resolved imaging, both for fundamental research and for applications in information technology. Transmission electron microscopy is a powerful technique for investigating the atomic structure, chemical composition and electromagnetic properties of materials with high spatial resolution and precision. However, the extraction of information about dynamic processes in the ps time regime is often not possible without extensive modification to the instrument while requiring careful control of the operation conditions to not compromise the beam quality. Here, we avoid these drawbacks by combining a delay line detector with continuous illumination in a transmission electron microscope. We visualize the gyration of a magnetic vortex core in real space and show that magnetization dynamics up to frequencies of 2.3 GHz can be resolved with down to ∼122ps temporal resolution by studying the interaction of an electron beam with a microwave magnetic field. In the future, this approach promises to provide access to resonant dynamics by combining high spatial resolution with sub-ns temporal resolution.

Quantitative measurement of charge accumulation along a quasi-one-dimensional W5O14 nanowire during electron field emission.

We use an electron holographic method to determine the charge distribution along a quasi-one-dimensional W5O14 nanowire during in situ field emission in a transmission electron microscope. The results show that the continuous charge distribution along the nanowire is not linear, but that there is an additional accumulation of charge at its apex. An analytical expression for this additional contribution to the charge distribution is proposed and its effect on the field enhancement factor and emission current is discussed.

Single Electron Precision in the Measurement of Charge Distributions on Electrically Biased Graphene Nanotips Using Electron Holography.

We use off-axis electron holography to measure the electrostatic charge density distributions on graphene-based nanogap devices that have thicknesses of between 1 and 10 monolayers and separations of between 8 and 58 nm with a precision of better than a single unit charge. Our experimental measurements, which are compared with finite element simulations, show that wider graphene tips, which have thicknesses of a single monolayer at their ends, exhibit charge accumulation along their edges. The results are relevant for both fundamental research on graphene electrostatics and applications of graphene nanogaps to single nucleotide detection in DNA sequencing, single molecule electronics, plasmonic antennae, and cold field emission sources.

Tunable Ampere phase plate for low dose imaging of biomolecular complexes.

A novel device that can be used as a tunable support-free phase plate for transmission electron microscopy of weakly scattering specimens is described. The device relies on the generation of a controlled phase shift by the magnetic field of a segment of current-carrying wire that is oriented parallel or antiparallel to the electron beam. The validity of the concept is established using both experimental electron holographic measurements and a theoretical model based on Ampere's law. Computer simulations are used to illustrate the resulting contrast enhancement for studies of biological cells and macromolecules.

Automated discrete electron tomography - Towards routine high-fidelity reconstruction of nanomaterials.

Electron tomography is an essential imaging technique for the investigation of morphology and 3D structure of nanomaterials. This method, however, suffers from well-known missing wedge artifacts due to a restricted tilt range, which limits the objectiveness, repeatability and efficiency of quantitative structural analysis. Discrete tomography represents one of the promising reconstruction techniques for materials science, potentially capable of delivering higher fidelity reconstructions by exploiting the prior knowledge of the limited number of material compositions in a specimen. However, the application of discrete tomography to practical datasets remains a difficult task due to the underlying challenging mathematical problem. In practice, it is often hard to obtain consistent reconstructions from experimental datasets. In addition, numerous parameters need to be tuned manually, which can lead to bias and non-repeatability. In this paper, we present the application of a new iterative reconstruction technique, named TVR-DART, for discrete electron tomography. The technique is capable of consistently delivering reconstructions with significantly reduced missing wedge artifacts for a variety of challenging data and imaging conditions, and can automatically estimate its key parameters. We describe the principles of the technique and apply it to datasets from three different types of samples acquired under diverse imaging modes. By further reducing the available tilt range and number of projections, we show that the proposed technique can still produce consistent reconstructions with minimized missing wedge artifacts. This new development promises to provide the electron microscopy community with an easy-to-use and robust tool for high-fidelity 3D characterization of nanomaterials.

Prospects for quantitative and time-resolved double and continuous exposure off-axis electron holography.

The technique of double exposure electron holography, which is based on the superposition of two off-axis electron holograms, was originally introduced before the availability of digital image processing to allow differences between electron-optical phases encoded in two electron holograms to be visualised directly without the need for holographic reconstruction. Here, we review the original method and show how it can now be extended to permit quantitative studies of phase shifts that oscillate in time. We begin with a description of the theory of off-axis electron hologram formation for a time-dependent electron wave that results from the excitation of a specimen using an external stimulus with a square, sinusoidal, triangular or other temporal dependence. We refer to the more general method as continuous exposure electron holography, present preliminary experimental measurements and discuss how the technique can be used to image electrostatic potentials and magnetic fields during high frequency switching experiments.

Dense, Regular GaAs Nanowire Arrays by Catalyst-Free Vapor Phase Epitaxy for Light Harvesting.

Density dependent growth and optical properties of periodic arrays of GaAs nanowires (NWs) by fast selective area growth MOVPE are investigated. As the period of the arrays is decreased from 500 nm down to 100 nm, a volume growth enhancement by a factor of up to four compared with the growth of a planar layer is observed. This increase is explained as resulting from increased collection of precursors on the side walls of the nanowires due to the gas flow redistribution in the space between the NWs. Normal spectral reflectance of the arrays is strongly reduced compared with a flat substrate surface in all fabricated arrays. Electromagnetic modeling reveals that this reduction is caused by antireflective action of the nanowire arrays and nanowire-diameter dependent light absorption. Irrespective of the periodicity and diameter, Raman scattering and grazing angle X-ray diffraction show signal from zinc blende and wurtzite phases, the latter originating from stacking faults as observed by high resolution transmission electron microscopy. Raman spectra contain intense surface phonons peaks, whose intensity depends strongly on the nanowire diameters as a result of potential structural changes and as well as variations of optical field distribution in the nanowires.

Rapid low dose electron tomography using a direct electron detection camera.

We demonstrate the ability to record a tomographic tilt series containing 3487 images in only 3.5 s by using a direct electron detector in a transmission electron microscope. The electron dose is lower by at least one order of magnitude when compared with that used to record a conventional tilt series of fewer than 100 images in 15-60 minutes and the overall signal-to-noise ratio is greater than 4. Our results, which are illustrated for an inorganic nanotube, are important for ultra-low-dose electron tomography of electron-beam-sensitive specimens and real-time dynamic electron tomography of nanoscale objects with sub-ms temporal resolution.

Tunable caustic phenomena in electron wavefields.

Novel caustic phenomena, which contain fold, butterfly and elliptic umbilic catastrophes, are observed in defocused images of two approximately collinear oppositely biased metallic tips in a transmission electron microscope. The observed patterns depend sensitively on defocus, on the applied voltage between the tips and on their separation and lateral offset. Their main features are interpreted on the basis of a projected electrostatic potential model for the electron-optical phase shift.