Limitations of beam damage in electron spectroscopic tomography of embedded cells

Elemental mapping in the energy filtering transmission electron microscope (EFTEM) can be extended into three dimensions (3D) by acquiring a series of two‐dimensional (2D) core‐edge images from a specimen oriented over a range of tilt angles, and then reconstructing the volume using tomographic methods. EFTEM has been applied to imaging the distribution of biological molecules in 2D, e.g. nucleic acid and protein, in sections of plastic‐embedded cells, but no systematic study has been undertaken to assess the extent to which beam damage limits the available information in 3D. To address this question, 2D elemental maps of phosphorus and nitrogen were acquired from unstained sections of plastic‐embedded isolated mouse thymocytes. The variation in elemental composition, residual specimen mass and changes in the specimen morphology were measured as a function of electron dose. Whereas 40% of the total specimen mass was lost at doses above 10⁶ e^?/nm², no significant loss of phosphorus or nitrogen was observed for doses as high as 10⁸ e^?/nm². The oxygen content decreased from 25 ± 2 to 9 ± 2 atomic percent at an electron dose of 10⁴ e^?/nm², which accounted for a major component of the total mass loss. The specimen thickness decreased by 50% after a dose of 10⁸ e^?/nm², and a lateral shrinkage of 9.5 ± 2.0% occurred from 2 × 10⁴ to 10⁸ e^?/nm². At doses above 10⁷ e^?/nm², damage could be observed in the bright field as well in the core edge images, which is attributed to further loss of oxygen and carbon atoms. Despite these artefacts, electron tomograms obtained from high‐pressure frozen and freeze‐substituted sections of C. elegans showed that it is feasible to obtain useful 3D phosphorus and nitrogen maps, and thus to reveal quantitative information about the subcellular distributions of nucleic acids and proteins.     

Tribological Characteristics of Nitrogen (N+) Implanted Iron

The effect of implantation of nitrogen ions (1.5 MeV) on the friction and wear characteristics of pure iron sliding against M-50 steel (unimplanted) was studied in a pin-on-disk sliding friction apparatus. Test conditions included room temperature (～25°C), a dry air atmosphere, a load of ½ kg (4.9 N), sliding velocities of 0.043 to 0.078 m/s (～15 to 25 rpm), a pure hydrocarbon lubricant (n-hexadecane), or a USP mineral oil and nitrogen ion implantation doses of 5 × 10¹⁵ and 5 × 10¹⁷ ions/cm².

No differences in wear rates were observed in the low-dose (5 × 10¹⁵ ions/cm²) experiments. In the high-dose experiments (5 × 10¹⁷ ions/cm²), small reductions in initial (～40 percent) and steady-state (～20 percent) wear rates were observed for nitrogen-implanted iron riders as compared with unimplanted controls. No differences in average friction coefficients were noted for either dose.

Auger electron spectroscopy combined with argon ion bombardment revealed a subsurface Gaussian nitrogen distribution with a maximum concentration of 6 atomic percent at a depth of 8 × 10^?7 m (0.8 μm). Similar analysis within the wear scar (～2.0 × 10^?5 m subsurface) of an implanted rider after 20 μm of wear yielded only background nitrogen concentration. No inward migration of nitrogen ions was observed.     

Optical transition process in AgOx super-resolution near-field structure

A sandwiched 15 nm AgO_x thin film of the super‐resolution, near‐field optical disk was studied using a confocal Z‐scan system. Nonlinear optical properties of quartz glass/ZnS–SiO₂ (170 nm)/AgO_x (15 nm)/ZnS–SiO₂ (40 nm) were measured using a Q‐switch Nd : YAG pulse laser of wavelength 532 nm, pulse width 0.7 ns, and 15.79 kHz repetition rate. Transmittance and reflectance of the sandwiched AgO_x thin film show important optical responses at the focused position of Z‐scan. The dissociation processes of AgO_x, recombination of the silver and oxygen, and the resonance of the localized surface plasmon of the nano‐composites of the AgO_x thin film are correlated to transmittance and reflectance at the focused position of the Z‐scan for different input laser powers. An irreversible upper threshold intensity of 4.40 × 10⁶ mW cm^?2 at the focused position was found. A reversible working window of the focusing intensity between 1.86 × 10⁶ and 4.40 × 10⁶ mW cm^?2 was measured with sandwiched AgO_x thin film alone. The near‐field interactions of the AgO_x thin film and the recording layers of super‐resolution near‐field optical disk are also discussed.     

Tem Of Dislocations Under High Stress In Germanium And Doped Silicon

The stacking fault energies y of silicon (58 ± 6 mJ m^?2) and germanium (75 ± 10 mJ m^?2) were determined. Within the limits of accuracy γ was not found to change on doping with (13·8 mol m^?3 (8 × 10¹⁸ cm^?3) boron, and 1·17 mol m^?3 (7 × 10¹⁷ cm^?3) phosphorus). Freezing in dislocations under high shear stress reveals a different behaviour of screw dislocations: whereas these dislocations become wider in pure and p-silicon, they become narrower in n-silicon. From this we conclude the ratio of mobilities of the two 30° partials to be different in n- and p-silicon. Other observations on frozen dislocations are mentioned.     

Parallel recording of electron energy loss spectra

Two silicon photo diode array devices were tested as parallel recording detectors for electron energy loss spectrometry (EELS). The direct bombardment of a Reticon photodiode array detector with high energy electrons (80 keV) causes an irreversible increase in diode dark current. The dark current saturates the detector amplifier after a dose of 10^?6 C/diode making it unsuitable for EELS. A scintillator coupled SIT vidicon is sensitive enough to count two high energy electrons with a spatial resolution of 100 μm, corresponding to 5 eV energy resolution with the electron optical system described. The large pixel-to-pixel gain variation inherent in the scintillator and vidicon can be reduced by averaging the spectrum over a large area of the target perpendicular to the dispersion direction. The L-edge of calcium for a 4 × 10^?3 weight fraction concentration biological specimen is observable in a 40 s parallel recorded spectrum. The minimum detectable concentration of calcium is estimated tobe ten times better for EELS than EDS X-ray analysis.     

Water loss at normal enamel histological points during air drying at room temperature

This in vitro study aimed to quantify water loss at histological points in ground sections of normal enamel during air drying at room temperature (25°C) and relative humidity of 50%. From each of 10 ground sections of erupted permanent human normal enamel, three histological points (n = 30) located at 100, 300 and 500 μm from enamel surface and along a transversal following prisms paths were characterized regarding the mineral, organic and water volumes. Water loss during air drying was from 0 to 48 h. Drying occurred with both falling and constant‐drying rates, and drying stabilization times (T_eq) ranged from 0.5 to 11 h with a mean 0.26 (±0.12)% weight loss. In some samples (n = 5; 15 points), T_eq increased as a function of the distance from the enamel surface, and drying occurred at an apparent diffusion rate of 3.47 × 10^?8 cm² s^?1. Our data provide evidence of air drying resulting in air replacing enamel's loosely bound water in prisms sheaths following a unidirectional water diffusion rate of 3.47 × 10^?8 cm² s^?1 (from the original enamel surface inward), not necessarily resulting in water evaporating directly into air, with important implications for transport processes and optical and mechanical properties.     

Evaluation of the effective cross‐sections for recombination and trapping in the case of pure spinel

In this paper, we have investigated the evolution of the secondary electron emission in the case of pure spinel during electron irradiation, achieved in a scanning electron microscope at room temperature, which is derived from the measurement of the induced and the secondary electron currents. It was observed from the experimental results, that there are two regimes during the charging process: a plateau followed by a linear variation, which are better identified by plotting the logarithm of the secondary electron emission yield lnσ as function of the total surface density of trapped charges in the material Q_T. For positive charging, E₀ = 1.1 and 5 keV, the slope of the linear part, whose value is of about 10^?10 cm² charge^?1, is independent of the primary electron energy. It is interpreted as a microscopic cross section for electron–hole recombination. For negative charging of pure spinel, E₀ = 15 and 30 keV, the slope is associated with an electron trapping cross section close to 10^?14 cm² charge^?1, which can be assigned to the microscopic cross section for electron trapping. This trapping cross section is four orders of magnitude lower than the recombination one.     

Electrochemical determination of hydrogen peroxide using a novel prussian blue–polythiophene–graphene oxide membrane-modified glassy carbon electrode

A polythiophene–graphene oxide compound membrane and Prussian blue were deposited sequentially on the surface of a glassy carbon electrode by cyclic voltammetry. Due to its excellent electrocatalysis and its analogy with peroxidase enzymes, Prussian blue has been widely used in amperometric biosensors. The polythiophene–graphene oxide compound membrane exhibited good electroconductibility and a large specific surface area. The fabricated Prussian blue/polythiophene/graphene oxide/glassy carbon electrode was characterized by transmission electron microscopy, scanning electron microscopy, and cyclic voltammetry. Under the optimal experimental conditions, the detection of hydrogen peroxide was studied by its amperometric current–time curve. Due to the presence of polythiophene–graphene oxide compound membrane and Prussian blue, the hydrogen peroxide sensor shows a linear calibration range of 1.0?×?10^?6–1.0?×?10^?4?mol?L^?1, detection limits of 3.2?×?10^?7?mol?L^?1 at a signal-to-noise ratio of 3, and recoveries from 95.0 to 105.0%. The results show that the modified glassy carbon electrode has potential practical application for the determination of hydrogen peroxide based on its sensitivity and long-term stability.     

Progress in electron microscopic diagnostics: semi-quantitative determination of precipitable calcium in different cell types of the organ of Corti in the guinea-pig

Potassium antimonate was used to precipitate calcium in the cochlea of the guinea-pig. The distribution of the calcium antimonate precipitates was analysed by electron microscopy. The precipitate density was determined in different cell types in the organ of Corti by counting the number of calcium binding sites in a 10-μm² area. The size of the precipitates varied considerably, and thus the relative amount of the precipitable calcium was estimated only semi-quantitatively. As the prominent carbon signal is superimposed over the nearby small Ca²⁺ -edge signals, the combined signal of the antimony M_4,5-edge and the oxygen K-edge of the calcium antimonate salt formed was chosen for the semi-quantitative estimation. Images of the inelastically scattered electrons of the precipitates at ΔE = 570 eV were recorded by electron spectroscopic imaging. The area covered by the calcium precipitates within a given cell type was determined in different ultrathin sections of the same organ of Corti by an image processing system.     

Prediction of phonon and electron contributions to thermal conduction in doped silicon films

The modified electron-phonon interaction model is proposed, and the relative contributions of phonons and electrons to the thermal conduction in doped silicon films are investigated quantitatively. The carrier contributions in sub-micro scale heat conduction are validated by an order of magnitude analysis and comparison with previous data. The simulation results show that the electron contribution in Si-layer is not negligible when the doping concentration is higher than 10¹⁹ cm^?3, which can be classified as semimetal or metal by the value of its electrical resistivity at 300 K. The electron thermal conductivity increases with increasing doping density, and its fraction is about 57.2% of the total thermal conductivity when the doping concentration is 5.0 × 10²⁰ cm^?3 and silicon film thickness 100 nm.     

Growing HgTe/Cd<Subscript>0.735</Subscript>Hg<Subscript>0.265</Subscript>Te quantum wells by molecular beam epitaxy

HgTe/Cd_0.735Hg_0.265Te nanostructures with HgTe quantum wells 16.2 and 21.0 nm thick are grown without additional doping on (013)CdTe/ZnTe/GaAs substrates by the method of molecular beam epitaxy. The compositions and thicknesses of the wide-gap layer and quantum well in the course of growth are performed by means of ellipsometry. The accuracy is Δx ? ±0.002 mole fractions of cadmium telluride in determining the composition and Δd ? 0.5 nm in determining the thickness of the wide-gap layer and quantum well. The central fragments of the wide-gap layers ≈ 10 nm thick are additionally doped by indium for a ～ 10¹⁵ cm^?3 volume concentration of charge carriers to be reached. Galvanomagnetic research in a wide range of magnetic field intensities at liquid helium temperatures reveals dimensional quantization levels and the presence of a two-dimensional electron gas in grown nanostructures. High mobility of the two-dimensional electron gas μ _e is obtained: 2 · 10⁵ and 5 · 10⁵ cm²/V · s for electron densities N _s equal to 1.5 · 10¹¹ and 3.5 · 10¹¹ cm^?2, respectively.     

Immersion‐Angle Dependence of the Resonant‐Frequency Shift of a Quartz Crystal Microbalance in Three Types of Newtonian Liquids

Abstract

The present work reports the new characteristics of the immersion‐angle dependence of the resonant‐frequency shift (ΔF) of the one‐face sealed quartz crystal microbalance (QCM) in three types of Newtonian liquids, i.e., sucrose, glucose, and glycerol solutions. Below some 1.80×10^?2 g cm^?2 · s^?1/2, the ΔF values are dependent on the immersion angles in all solutions. However, we have found that the transition phenomenon of ΔF occurs between 1.78×10^?2 and 4.80×10^?2 g · cm^?2 · s^?1/2 in the sucrose solution, between 1.75×10^?2 and 4.34×10^?2 g · cm^?2 · s^?1/2 in the glucose solution and between 1.83×10^?2 and 3.03×10^?2 g · cm^?2 · s^?1/2 in the glycerol solution, respectively. Moreover, above the concentrations of the end points of the transition phenomenon, the ΔF values of the sucrose solution are equal to those of 90° at all immersion angles. On the other hand, those of the glucose and the glycerol solutions are the same as those of 30°. This difference may be caused by inherent characteristics of adsorption to the surface of the QCM electrode.     

A monitor for heavy ion flux density control based on detection of recoil protons during irradiation of polymer films

An monitor for control of accelerated particle flux density used in irradiation of polymer films with heavy ions with a low track density (10³?5 × 10⁷ cm^?2) is based on detection of recoil protons. It has been designed to control irradiation of a film with a width of up to 650 mm and is composed of five PIN diodes with a hydrogenous target placed in front of each diode. The sensitivity of the monitor with a polyethylene target to the density of a xenon ion beam is described by a linear function; its value is (4.0±0.5) × 10^?3 pulses/(ion/cm²). The instrument is capable of monitoring ion flux densities of <10⁶ cm^?2, for which all other methods are ineffective. It can be used to determine the track density in a finished product, bypassing the stage of a check with an electron microscope. The signals from the monitor can also be used to introduce a feedback in a system of automatic beam tuning in order to improve the film irradiation quality. The radiation hardness of silicon detectors is ≈ 10¹² protons/cm². At an average detector load of ≈ 10⁴ protons/cm², the average service life of the monitor is 10⁸ s of continuous operation.     

Hydrothermal synthesis of a lamellar zinc dimolybdate hydroxide with application as the anode for lithium-ion batteries

Abstract

A lamellar zinc dimolybdate hydroxide Zn₃Mo₂O₈(OH)₂ was prepared for the first time by a facile hydrothermal synthesis. The electrochemical properties of the lamellar Zn₃Mo₂O₈(OH)₂ as an anode material were investigated by cyclic voltammetry (CV), reversible capacity, cycling stability and rate capability for lithium ion batteries (LIB). The lamellar zinc dimolybdate hydroxide exhibits a reversible capacity of 404.6 mAh g^?1 at a current density of 100?mA g^?1 after 200 cycles. The reversible capacity of the lamellar Zn₃Mo₂O₈(OH)₂ remained at 60 mAh g^?1 even at a current density of 3000?mA g^?1. When the current density was returned to 100?mA g^?1, a discharge capacity of 380 mAh g^?1 was maintained after 200 discharge/charge cycles. The excellent electrochemical performance may be due to its unique lamellar structure, which buffers the volume change during the Li⁺ intercalation/de-intercalation and provides the electrode with convenient lithium ions and electron transport pathways. These results suggest the promising potential application of the lamellar zinc dimolybdate hydroxide in lithium-ion batteries.     

Morphological studies of pseudowollastonite for biomedical application

Pseudowollastonite ceramic (psW) composed of CaO·SiO₂ was found to be bioactive in a simulated body fluid environment. The chemical reaction initiated at the material surface resulted in hydroxyapatite (HA)formation. These bone-bonding properties are essential for securing the necessary physico-chemical integration of the material with living bone. Materials behaving in this way can be considered for potential biomedical application as bone tissue substitute for a natural bone repair or replacement as implant. A mechanism of hydroxyapatite formation on pseudowollastonite ceramics surface was investigated during exposure to a simulated body fluid (SBF) for a period of 3 weeks. Morphology and structure of the surface product and its original substrate was examined by thin-film X-ray diffraction, scanning electron microscopy and high-resolution transmission electron microscopy. HA crystals were found to form on an amorphous silica intermediate layer. (100) lattice planes of HA were resolved and identified. Concentration of ions in the SBF and pH of the SBF were monitored throughout the exposure. Additional pH measurements were made at the interface of psW with SBF. The HA formation occurred when there was a sudden increase of pH from 7·25 to 10·5 at the interface of psW with SBF as a result of ionic exchange between 2H⁺ and Ca²⁺ within the psW network. This ionic exchange transformed the psW crystals into an amorphous silica phase. The appropriate pH and the ion concentrations were essential for partial dissolution of the amorphous silica phase and subsequent precipitation of a Ca-P rich phase which then transformed to HA.     

Phase separation as a basis for the formation of light-emitting silicon nanoclusters in SiO x films irradiated with swift heavy ions

This paper presents a study of the effect of swift heavy Xe ions of energy 130–167 MeV at doses of 10¹²–10¹⁴ cm^?2 and Bi ions of 700 MeV at doses of 3·10¹²–3·10¹³ cm^?2 on films of stoichiometric thermal silicon dioxide, silicon dioxide films with ion-implanted excess silicon, and SiO _x films with the stoichiometric parameter x varying from 0 to 2. According to electron microscopy and Raman spectroscopy data, irradiation with the swift heavy ions resulted in the formation of silicon nanoclusters. The luminescence spectra depended on the size, number, and structure of the Si nanoclusters formed. Their size can be controlled by varying both the effect parameters (primarily, the ion energy loss per unit length of the track) and the stoichiometric composition of the films.     

An infrared interferometer for investigating subthermonuclear plasma in the GOL-3 multimirror trap

A simple Michelson CO₂ interferometer for measuring the high-temperature plasma density within one interference fringe (n _e l = 10¹⁶ cm^?2) at two points (0.8 and 9.0 m) of the long (L = 12 m) GOL-3 corrugated trap was used. A piezoelectric element that provides displacements of the mirror in the reference arm of the interferometer is used to calibrate the interferometer and perform remote control of the initial measurement phase. The interferometer is manufactured from dielectric materials, thus excluding a mechanical action of stray magnetic fields on its elements. The time resolution of the interferometer is determined by a HgCdTe diode and equals ??1 ns. The sensitivity of the interferometer is ??5 × 10^?4 of a fringe (n _e l = 2 × 10¹³ cm^?2).     

Data recording system of a dispersion interferometer based on a CO<Subscript>2</Subscript> laser

The data recording system of a multichannel double-pass dispersion interferometer based on a CO₂ laser is described. This system has been designed to record the linear density of plasmas in a real-time mode with a time discreteness of 4 μs and resolution 〈N _e L〉 ～ 0.34 × 10¹³ cm^?2 (N _e is the electron component of the plasma density, and L is the plasma size in the wave propagation direction) in the range of linear density variations of up to 10¹⁷ cm^?2. The system is built from unified recording modules that use fast ADCs to record the shape of photodetector and modulator signals and FPGA-based digital units of dataflow processing to form results of measurements. The single-channel recording module of the dispersion interferometer has been tested under actual experimental conditions of the GDL gas-dynamic trap and the TEXTOR tokamak (Julich, Germany).     

Contrast effects in photoelectron microscopy; UV dose-dependent quantum yields of biological surface components

The relative brightness of photoelectron microscopy images as a function of exposure to UV light has been determined from model systems representative of biological cell surface components. Quantitative data for amino acid homopolymers, L-α-dipalmitoylphosphatidylcholine, and the polysaccharide Ficoll are reported as the absolute photoelectron quantum yields. The photoelectron quantum yields increase substantially over the initial values. For example, the quantum yields of poly-L-tyrosine at 200 nm is initially about 5 × 10^?8 electrons/incident photon. The quantum yield increases with 254 nm irradiation, leveling off at about 5 × 10^?4 electrons/incident photon after a dose of 3 × 10²¹ quanta cm^?2. Pre-irradiation of poly-L-tyrosine in the presence of certain chemical agents, for example, the Lewis base diborane (B₂H₆), results in a substantial reduction of the dose-dependent increase in quantum yield. Exposure to the reducing agent stannane (SnH₄) essentially eliminates the effect. These chemical treatments provide methods of controlling the UV dose-dependent effects in the photoelectron images.     

Material contrast in SEM: Fermi energy and work function effects

‘Is it possible to assign various grey levels of a scanning electron microscope (SEM) image to different components of a given sample? Among other instrumental effects, the answer is not only a function of the respective secondary electron emission (SEE) yields of the components, δ, but also of the angular fraction of the secondary electrons (SE)s being collected, k_α and of a possible voltage contact effect between sample and detector, k_?. Expressed as a function of E_F, Fermi energy, and ?, work function of the components of interest, equations of spectral, (∂δ/∂E_k), and angular, (∂δ/∂α) distributions of the emitted SEs permit to evaluate k_α and k_? for Au and Si. It has been established that collected SE spectra, ∂δ_α/∂E_k, are distorted with respect to the emitted and fraction k_α is material dependent for a solid angle of detection Ω° less than 2π (or maximum semi-apex angle α_max<90°) In particular, for coaxial detections around the normal incident beam the detected fraction of SEs from Au, k_α(Au), is slightly larger than that for Si, k_α(Si). For simple geometries in the vacuum gap, similar investigations show that parameter k_φ is also larger for gold than for n-doped Si as well as for p-doped Si with respect to n-doped Si. Then Au is always quite brighter than n-doped Si in the SEM images while a doping contrast, C, due to a work function effect may reach ∼15% for a Si p/n junction with N_p∼10¹⁶ and N_n∼10¹⁵ cm⁻³. The present analysis may be extended to some metals such as Ag, Cu, Pb, Pd, Pt, and Zn that are expected to appear brighter than Si(n) and Ge in the SEM images.