High-resolution Electron Image Research Articles

SigmaR1 shapes rough endoplasmic reticulum membrane sheets

Rough endoplasmic reticulum (ER) sheets are a fundamental domain of the ER and the gateway into the secretory pathway. Although reticulon proteins stabilize high-curvature ER tubules, it is unclear whether other proteins scaffold the flat membranes of rough ER sheets. Through a proteomics screen using ER sheet-localized RNA-binding proteins as bait, we identify the sigma-1 receptor (SigmaR1) as an ER sheet-shaping factor. High-resolution live cell imaging and electron tomography assign SigmaR1 as an ER sheet-localized factor whose levels determine the amount of rough ER sheets in cells. Structure-guided mutagenesis and invitro reconstitution on giant unilamellar vesicles further support a mechanism whereby SigmaR1 oligomers use their extended arrays of amphipathic helices to bind and flatten the lumenal leaflet of ER membranes to oppose membrane curvature and stabilize rough ER sheets.

High-resolution multi-mode electron and ion imaging spectrometer at SXFEL

A new multi-mode electron and ion (MEI) imaging spectrometer with two arms of VMI and COLTRIMS/VMI (velocity map imaging/cold-target recoil-ion momentum spectroscopy) is designed to combine various photoelectron and ion detection modes for experiments at Shanghai soft x-ray free-electron laser (SXFEL) facility. The experiments can be optionally operated either with both ion and electron detection in a coincidence/covariance manner (VMI arm and COLTRIMS/VMI arm), or only photoelectron/photoion is detected with the high-resolution VMI arm. The simulated resolutions for 30–150 eV photoelectrons and 3.3 eV–18.0 eV N+ and N2+ photoions are up to 1.0% and 3.1%–1.0% according to our simulation, respectively. MEI spectrometer is expected to improve the experimental abilities significantly considering the low-repetition rate of the SXFEL and to enable the investigation of a diverse range of atomic and molecular phenomena triggered by soft x-ray free electron laser irradiation.

Whole-Cell Electron Tomography Analysis of Vacuole Biogenesis.

Vacuoles in plant cells are the most prominent organelles that harbor distinctive features, including lytic function, storage of proteins and sugars, balance of cell volume, and defense responses. Despite their dominant size and functional versatility, the nature and biogenesis of vacuoles in plants per se remain elusive and several models have been proposed. Recently, we used the whole-cell 3D electron tomography (ET) technique to study vacuole formation and distribution at nanometer resolution and demonstrated that small vacuoles are derived from multivesicular body maturation and fusion. Good sample preparation is a critical step to get high-quality electron tomography images. In this chapter, we provide detailed sample preparation methods for high-resolution ET in Arabidopsis thaliana root cells, including high-pressure freezing, subsequent freeze-substitution fixation, embedding, and serial sectioning.

Synthesis of nanocomposites of montmorillonite with carbon nanotubes as a potential material for water purification

The main goal of this research is to create nanocomposites based on unmodified and iron-modified (FeNP) montmorillonite (Mt) and carbon nanotubes (CNT) synthesized using the chemical vapor deposition method. The target area for the application of these materials is the creation of water treatment systems. This paper compares the efficiency of the CNT synthesis process on Mt before and after modification with FeNP of different concentrations and provides the characterization of the CNT microstructure and structure using different methods, such as scanning electron microscopy, high-resolution transmission electron imaging, and Raman spectroscopy. For initial verification of properties important for water purification, Mt+CNT and Mt+FeNP+CNT nanocomposites on a carbon nonwoven fabric (CF) are tested in this work. Incubation of the above-mentioned samples in a water–oil mixture reveals complex adsorption dynamics. The CF+Mt+FeNP+CNT sample shows a very good oil adsorption capacity due to its superhydrophobic and oleophilic properties.

Targeting of membrane proteins with fluoronanogold probes for high-resolution correlative microscopy.

Correlative light and electron microscopy (CLEM) can provide valuable information about a biological sample by giving information on the specific localization of a molecule of interest within an ultrastructural context. In this work, we describe a simple CLEM method to obtain high-resolution images of neurotransmitter receptor distribution in synapses by electron microscopy (EM). We use hippocampal organotypic slices from a previously reported mouse model expressing a modified AMPA receptor (AMPAR) subunit that binds biotin at the surface (Getz et al., 2022). This tag can be recognized by StreptAvidin-Fluoronanogold™ conjugates (SA-FNG), which reach receptors at synapses (synaptic cleft is 50-100nm thick). By using pre-embedding labeling, we found that SA-FNG reliably bind synaptic receptors and penetrate around 10-15μm in depth in live tissue. However, the silver enhancement was only reaching the surface of the slices. We show that permeabilization with triton is highly effective at increasing the in depth-gold amplification and that the membrane integrity is well preserved. Finally, we also apply high-resolution electron tomography, thus providing important information about the 3D organization of surface AMPA receptors in synapses at the nanoscale.

Ag-Doped Free-Standing 2D TiO2 Sheets: Electronic, Optical, Magnetic, and Self-Healing Behaviour.

Beyond a critical doping level, Ag - 2D TiO2 sheets (ATO) are deemed to be a flexible transparent conductor, useful for visible-range functional photonic/optoelectronic devices/sensors, sunlight-sensitive catalysis, and light-activated resistive switching. Due to the lack of control of surface energy which often leads to the formation of structural defects and even dimensionality crossover (2D to 0D) of materials during doping reaction, it is challenging to obtain ATO with a controlled doping level. Gauging the urgency, therefore we report the surface energy-controlled synthesis of ATO employing liquid phase exfoliation of TiO2 and subsequent hydrothermal Ag-doping in the presence of Hexamethylenetetramine (HMTA). Electron microscopy and atomic force microscopy reveal ATO sheets with large lateral dimensions. 6-fold, 4-fold, and strain-mediated crystallographic phases of 2D ATO have been revealed by high-resolution electron imaging. Successful tuning of the band gap down to ~ 2 eV with Ag doping up to ~ 10% is obtained. Synthesized 2D ATO have been investigated for their electrical, optical, optoelectronic, photoluminescence, and ferromagnetic behaviour. Visible light-sensitive thermally/structurally robust semiconductor/conductor via tuneable doping will pave the way for their flexible as well as wearable device applications. Self-healing effect of AFM tip-generated mechanical stress has also been demonstrated.

Lossless Monochromator in an Ultrafast Electron Microscope Using Near-Field THz Radiation.

The ability to form monoenergetic electron beams is vital for high-resolution electron spectroscopy and imaging. Such capabilities are commonly achieved using an electron monochromator, which energy filters a dispersed electron beam, thus reducing the electron flux to yield down to meV energy resolution. This reduction in flux hinders the use of monochromators in many applications, such as ultrafast transmission electron microscopes (UTEMs). Here, we develop and demonstrate a mechanism for electron energy monochromation that does not reduce the flux-a lossless monochromator. The mechanism is based on the interaction of free-electron pulses with single-cycle THz near fields, created by nonlinear conversion of an optical laser pulse near the electron beam path inside a UTEM. Our experiment reduces the electron energy spread by a factor of up to 2.9 without compromising the beam flux. Moreover, as the electron-THz interaction takes place over an extended region of many tens of microns in free space, the realized technique is highly robust-granting uniform monochromation over a wide area, larger than the electron beam diameter. We further demonstrate the wide tunability of our method by monochromating the electron beam at multiple primary electron energies from 60 to 200keV, studying the effect of various electron and THz parameters on its performance. Our findings have direct applications in the fast-growing field of ultrafast electron microscopy, allowing time- and energy-resolved studies of exciton physics, phononic vibrational resonances, charge transport effects, and optical excitations in the mid IR to the far IR.

Scrutinizing pre- and post-device fabrication properties of atomic layer deposition WS2 thin films

In this work, we investigate the physical and electrical properties of WS2 thin films grown by a plasma-enhanced atomic layer deposition process, both before and after device fabrication. The WS2 films were deposited on thermally oxidized silicon substrates using the W(NMe2)2(NtBu)2 precursor and a H2S plasma at 450 °C. The WS2 films were approximately 8 nm thick, measured from high-resolution cross-sectional transmission electron imaging, and generally exhibited the desired horizontal basal-plane orientation of the WS2 layers to the SiO2 surface. Hall analysis revealed a p-type behavior with a carrier concentration of 1.31 × 1017 cm−3. Temperature-dependent electrical analysis of circular transfer length method test structures, with Ni/Au contacts, yielded the activation energy (Ea) of both the specific contact resistivity and the WS2 resistivity as 100 and 91 meV, respectively. The similarity of these two values indicates that the characteristics of both are dominated by the temperature dependence of the WS2 hole concentration. Change in the material, such as in sheet resistance, due to device fabrication is attributed to the chemicals and thermal treatments associated with resist spinning and baking, ambient and UV exposure, metal deposition, and metal lift off for contact pad formation.

Interface tomography of GaInAs/AlInAs quantum cascade laser active regions

We present a high-resolution electron tomography study of buried ultra-thin layers and their interfaces from the active region of a (Ga,In)As/(Al,In)As quantum cascade laser (QCL) test structure. Using a high-angle annular dark-field scanning transmission electron microscopy image series, a three-dimensional (3D) reconstruction of the complex layer structure is obtained. From this 3D information, we determine quantitative values for the chemical width and, simultaneously and independently, the root mean square roughness (rms) and lateral correlation length of the individual interfaces of a cascade using topographic height maps. The interfacial widths are comparably small for all interfaces within a cascade and the layer thicknesses show only a small standard deviation of less than one monolayer. The rms roughness is systematically lower at the direct (Ga,In)As-on-(Al,In)As interface compared to its inverse. In addition, using the one-dimensional height–height correlation function, different lateral correlation lengths along the two perpendicular in-plane directions are detected indicating a distinct anisotropy of the interface morphology. Our accurate and comprehensive results on the QCL test structure will serve as feedback to evaluate the growth process and help to assess the performance of corresponding future laser devices in detail.

Wafer-Scale Fabrication of Hierarchically Porous Silicon and Silica by Active Nanoparticle-Assisted Chemical Etching and Pseudomorphic Thermal Oxidation.

Many biological materials exhibit a multiscale porosity with small, mostly nanoscale pores as well as large, macroscopic capillaries to simultaneously achieve optimized mass transport capabilities and lightweight structures with large inner surfaces. Realizing such a hierarchical porosity in artificial materials necessitates often sophisticated and expensive top-down processing that limits scalability. Here, an approach that combines self-organized porosity based on metal-assisted chemical etching (MACE) with photolithographically induced macroporosity for the synthesis of single-crystalline silicon with a bimodal pore-size distribution is presented, i.e., hexagonally arranged cylindrical macropores with 1 µm diameter separated by walls that are traversed by pores 60 nm across. The MACE process is mainly guided by a metal-catalyzed reduction-oxidation reaction, where silver nanoparticles (AgNPs) serve as the catalyst. In this process, the AgNPs act as self-propelled particles that are constantly removing silicon along their trajectories. High-resolution X-ray imaging and electron tomography reveal a resulting large open porosity and inner surface for potential applications in high-performance energy storage, harvesting and conversion or for on-chip sensorics and actuorics. Finally, the hierarchically porous silicon membranes can be transformed structure-conserving by thermal oxidation into hierarchically porous amorphous silica, a material that could be of particular interest for opto-fluidic and (bio-)photonic applications due to its multiscale artificialvascularization.

Resolving local microstructure variations in Zr-2.5Nb CANDU pressure tubes using high spatial resolution electron backscatter diffraction and imaging

The alloy Zr-2.5Nb is used for manufacturing the pressure tubes installed in the core of Canada Deuterium Uranium (CANDU) nuclear reactors. This alloy consists of α-zirconium with hexagonal close-packed (HCP) crystals and a minor β-zirconium phase with body centered cubic (BCC) crystals that are mainly located in between the α-grains. Due to the significant elastic and plastic anisotropy of HCP crystals as well as the development of strong textures during manufacturing, these tubes exhibit significant mechanical anisotropy. However, both local microstructure and texture can change, affecting the local patterning of stress fields and the subsequent hydrogen embrittlement rate. Resolving the local microstructure variation of Zr-2.5Nb is challenging due to the development of nano-sized grains during manufacturing. This paper focuses on developing a procedure for the consistent characterization of such local variations using high spatial resolution electron backscatter diffraction (EBSD) and imaging. Microstructure variations along the radial and transverse directions of the transverse-radial plane, as well as along the axial direction of the transverse-axial and radial-axial planes are studied. It is shown that microstructure variations are non-negligible. For the locations that the transverse-radial planes are studied, pole figures rotate, and Kearns factors decrease along the radial direction. In addition, the morphologies of both α- and β-crystals change significantly along the radial direction. With the procedure used for characterization, some β-crystals are indexed and their orientation relationship with their neighboring α-grains are examined.

Chemically coupling SnO2 quantum dots and MXene for efficient CO2 electroreduction to formate and Zn–CO2 battery

Electrochemical conversion of CO2 into formate is a promising strategy for mitigating the energy and environmental crisis, but simultaneously achieving high selectivity and activity of electrocatalysts remains challenging. Here, we report low-dimensional SnO2 quantum dots chemically coupled with ultrathin Ti3C2Tx MXene nanosheets (SnO2/MXene) that boost the CO2 conversion. The coupling structure is well visualized and verified by high-resolution electron tomography together with nanoscale scanning transmission X-ray microscopy and ptychography imaging. The catalyst achieves a large partial current density of -57.8 mA cm-2 and high Faradaic efficiency of 94% for formate formation. Additionally, the SnO2/MXene cathode shows excellent Zn-CO2 battery performance, with a maximum power density of 4.28 mW cm-2, an open-circuit voltage of 0.83 V, and superior rechargeability of 60 h. In situ X-ray absorption spectroscopy analysis and first-principles calculations reveal that this remarkable performance is attributed to the unique and stable structure of the SnO2/MXene, which can significantly reduce the reaction energy of CO2 hydrogenation to formate by increasing the surface coverage of adsorbed hydrogen.

The role of volatile organic compounds in the oxidation-driven fragmentation of soot particles

In this work, the role of volatile organic compounds (VOCs) in soot oxidation and fragmentation was investigated in a lean, premixed CH4 flame. An aerosol generator was used to homogeneously disperse soot with and without VOCs (termed Soot-W and Soot-WO, respectively) into the flame. Because minimal soot was formed in the flame itself, information on the fragmentation of the Soot-W and Soot-WO during the oxidation process was readily captured to determine the role of VOCs in soot fragmentation. The integrated number concentration of fine mode particles and the degree of particle aggregation show that the presence of VOCs suppresses soot aggregate fragmentation, primarily by the mitigating consumption of the neck sites between primary particles due to the decreased specific surface area. The actual oxygen diffusion route and effective pore width between the crystallite structures were determined using the image analysis technique. To determine the characteristic regime of oxygen diffusion, the distribution of effective pore width was studied using high-resolution transmission electron microscopy and image analysis. The estimated effectiveness factor indicates that the presence of VOCs lowers the propensity of internal burning via intra-particle oxygen diffusion, and reduces the extent of primary particle fragmentation, as evidenced by a relatively smaller peak of ultrafine particles. The analysis of the nanostructure of neck sites vs. the bulk discovers significantly more disordered and thus reactive structure at neck sites, explaining the prevalence of aggregate fragmentation over primary particle fragmentation. The finding of the role of VOCs in oxidation-driven fragmentation is beneficial to the optimization of existing models for predicting the number concentration of fine and ultrafine particles. Furthermore, a better understanding of this role may aid in the control of the most harmful fine and ultrafine particle emissions.

Influence of Global Operating Parameters on the Reactivity of Soot Particles from Direct Injection Gasoline Engines

The aim of this study is to investigate the impact of global operating parameters, e.g., engine speed, brake mean effective pressure, and air–fuel ratio, of a turbocharged 4-cylinder GDI engine on the reactivity of soot particles against oxidation. The knowledge of soot reactivity is crucial for optimizing gasoline particulate filter regeneration strategies and is, consequently, a key parameter for reducing fuel consumption and CO2 emissions. In this work, time-resolved in-cylinder soot concentrations and exhaust particle size distributions are measured by using two-color pyrometry, engine exhaust particle sizer and smoke meter, respectively. Reactivity against oxidation by molecular oxygen is determined by temperature programmed oxidation analysis. To derive a physicochemical explanation for varying soot reactivity, the morphological and nanostructural primary particle structure of collected samples is analyzed using high-resolution electron microscopy and image analysis algorithms. The results reveal that engine operating parameters affect soot reactivity differently. While engine speed has only a slight effect, a reduction of air/fuel ratio (λ < 1.0) or an increase of BMEP > 10 bar significantly reduces the soot oxidation reactivity. These findings give evidence, that the quality of the fuel/air mixture is a significant parameter influencing soot reactivity. Measured soot concentrations substantiate the hypothesis that low-sooty homogeneous premixed combustion of a homogeneous fuel/air mixture favors formation of high-reactive soot particle fractions. Reactive soot particle aggregates are composed of multiple soot fractions of different reactivity. Reactive primary particles are composed of short graphene-like layers and vice versa, providing a physicochemical explanation for varying soot reactivity depending on engine operating conditions.

3D Atomic Structure of Supported Metallic Nanoparticles Estimated from 2D ADF STEM Images: A Combination of Atom-Counting and a Local Minima Search Algorithm.

Determining the 3D atomic structure of nanoparticles (NPs) is critical to understand their structure-dependent properties. It is hereby important to perform such analyses under conditions relevant for the envisioned application. Here, the 3D structure of supported Au NPs at high temperature, which is of importance to understand their behavior during catalytic reactions, is investigated. To overcome limitations related to conventional high-resolution electron tomography at high temperature, 3D characterization of NPs with atomic resolution has been performed by applying atom-counting using atomic resolution annular dark-field scanning transmission electron microscopy (ADF STEM) images followed by structural relaxation. However, at high temperatures, thermal displacements, which affect the ADF STEM intensities, should be taken into account. Moreover, it is very likely that the structure of an NP investigated at elevated temperature deviates from a ground state configuration, which is difficult to determine using purely computational energy minimization approaches. In this paper, an optimized approach is therefore proposed using an iterative local minima search algorithm followed by molecular dynamics structural relaxation of candidate structures associated with each local minimum. In this manner, it becomes possible to investigate the 3D atomic structure of supported NPs, which may deviate from their ground stateconfiguration.

HRTEM observation of morphological and structural evolution of aromatic fringes during the transition from coal to graphite

The maximum reflectance of vitrinite or graphite microcrystals in Yongan Coalfield, Fujian Province, China, ranges from 5.33% to 10.21%, comprising meta-anthracite, semi-graphite, and graphite. The morphological characteristics of aromatic fringes during different evolution stages were quantitatively analyzed using high-resolution transmission electron microscopy and image processing techniques. The results show that a nonlinear evolution of aromatic fringe morphology in the process of coal graphitization is manifested in four aspects, involving the rapid connection and growth, enhancement of ordering alignment, increase of stacking and development of straightening. In meta-anthracite samples, 3 × 3 aromatic rings representing short aromatic fringes are dominated, a majority of aromatic fringes were curved, disjoint or separate, and show a turbostratic layers, with poorly aligned. The spacing between adjacent aromatic fringes exceeded the range of graphitized carbon, and structural defects, such as nanopores were generally visible. In semi-graphite samples, in addition to 3 × 3 aromatic rings, the proportion of >8 × 8 aromatic rings represented longer aromatic fringes increasing significantly, along with ordering alignment and straightening. Stacking with two more layers increased and the spacing between adjacent aromatic fringes decreased to the range of graphitized carbon, and the structural defects decreased considerably. In graphite sample, the proportion of >8 × 8 aromatic rings increase significantly, the length, stacking, and ordering alignment of aromatic fringes increased abruptly, the spacing between adjacent aromatic fringes remained within the range of natural graphite, and a rapid graphite-like structure with flat aromatic fringes formed. A stage evolution of aromatic fringe morphology, whereas remaining almost constant of mainly organic elements, which imply the evolution of aromatic fringes in meta-anthracite originated from ring condensation dominated by late coalification, graphitization starts at semi-graphite stage and enhanced during graphite stage. The making up, originating from linkup and pileup, considered as a special mechanism for the graphitization of aromatic fringes.

Ion-Induced Bending with Applications for High-Resolution Electron Imaging of Nanometer-Sized Samples

Ion-Induced Bending with Applications for High-Resolution Electron Imaging of Nanometer-Sized Samples

Nanoscale coating on tip geometry by cryogenic focused ion beam deposition

We report application of nanoscale in-situ cryo-condensation coating technique on needle-shaped specimens and frozen hydrated cells. By using a cryogenic focused ion beam/ scanning electron microscopy (cryo-FIB/SEM) instrument, phase transformation of the metal–organic precursor gas has been demonstrated to occur at the cryogenically cooled metal tip of 50 nm in radius, and the properties of the deposited layer can be tuned by the control parameters. Revealed surface morphologies on the needle tip include nanoscale columns using glancing angle deposition (GLAD). With deposition performed at multiple orientations, a continuous frozen shell as thin as 40 nm can be formed. The physical mechanisms of curing by ion beam/electron beam irradiation have been further investigated with characterization tools including transmission electron microscopy (TEM) and energy dispersive x-ray spectroscopy (EDS), and the results confirm the feasibility of constructing a platinum-rich conductive layer. The proposed approach is also demonstrated by depositing a thin conductive layer on frozen hydrated bacterial cells to facilitate sectioning and high-resolution electron imaging. The proposed approach opens a novel route to nanofabrication on tip geometry as well as site-specific conductive protection to facilitate nanocharacterisation.

High Pressure in Boron Nitride Nanotubes for Kirigami Nanoribbon Elaboration

Cutting and folding 2D systems is one of the explored paths to tune physical and chemical properties in one-atom-thick matter. Contrary to graphene, boron nitride (BN) nanoribbons are difficult to obtain, and folded BN nanoribbon structures have not been reported yet. Here, we show that pressure application in multiwalled boron nitride nanotubes leads to different types of tube internal organizations including BN nanoribbon formation and folds. The new structures are associated with the breaking of a number of the internal tubes, leading to either nonorganized structures in the form of internal tube alveoli or an organized stacking of folded h-BN nanoribbons. Irreversible changes in the morphology of multiwalled BN nanotubes (MWBNNTs) take place from ∼7 GPa, and morphologically modified tubes could be observed up to pressures of at least 49 GPa. The experimental probes utilized included high-resolution transmission microscopy, electron tomography, and Raman spectroscopy. Atomistic modeling shows the formation of pinch structures along the tubes that favor pressure-induced bond-breaking and hybridization changes and confirm the folded structure. Both experiments and modeling show that tube polygonization is a prominent characteristic of MWBNNTs even at ambient pressure. Overall, the pressure evolution of MWBNNTs strongly differs from their carbon analogues. The high mechanical stability of BN tube geometry is of interest for composite-based structural materials. On the other side, the availability of h-BN nanoribbons and folded structures opens new prospects to produce physically modified BN properties.

Hrtem Observation of Morphological and Structural Evolution of Aromatic Fringes During the Transition from Coal to Graphite

The maximum reflectance of vitrinite or graphite microcrystals in Yongan Coalfield, Fujian Province, China, ranges from 5.33% to 10.21%, comprising meta-anthracite, semi-graphite, and graphite. The morphological characteristics of aromatic fringes during different evolution stages were quantitatively analyzed using high-resolution transmission electron microscopy and image processing techniques. The results show that a nonlinear evolution of aromatic fringe morphology in the process of coal graphitization is manifested in four aspects, involves the rapid connection and growth, enhancement of ordering alignment, increase of stacking and development of straightening. The stage evolution of aromatic fringe morphology and remained almost constant of mainly organic elements, implying the evolution of aromatic fringes in meta-anthracite originated from ring condensation dominated by late coalification, graphitization starts at semi-graphite stage and enhanced during graphite stage. The making up, originating from linkup and pileup, considered as a special mechanism for the graphitization of aromatic fringes.