Simultaneous observation of multiple interferograms with Mach-Zehnder type electron interferometer on a 1.2-MV field-emission transmission electron microscope.

We developed a Mach-Zehnder type electron interferometer (MZ-EI) that enables simultaneous observation of interferograms created at multiple output locations on a 1.2-MV field-emission transmission electron microscope. This MZ-EI is composed of two single-crystal thin films, a lens located between the single-crystal thin films, and imaging lenses. By comparing interferograms created by electron waves travelling through different beam paths, we found that the relative phase difference was caused by phase modulation passing through the single crystals and by aberrations and defocus values of the lenses. We also confirmed that the relative phase difference can be controlled using the tilted illumination method.

Origin of giant enhancement of phase contrast in electron holography of modulation-doped n-type GaN.

The electron optical phase contrast probed by electron holography at n-n+ GaN doping steps is found to exhibit a giant enhancement, in sharp contrast to the always smaller than expected phase contrast reported for p-n junctions. We unravel the physical origin of the giant enhancement by combining off-axis electron holography data with self-consistent electrostatic potential calculations. The predominant contribution to the phase contrast is shown to arise from the doping dependent screening length of the surface Fermi-level pinning, which is induced by FIB-implanted carbon point defects below the outer amorphous shell. The contribution of the built-in potential is negligible for modulation doping and only relevant for large built-in potentials at e.g. p-n junctions. This work provides a quantitative approach to so-called dead layers at TEM lamellas.

Association between hepatitis C virus genotype 4 and renal cell carcinoma: Molecular and virological studies.

Hepatitis C virus (HCV) is the most common infection worldwide. The correlation between HCV and renal cell carcinoma (RCC) is still mysterious. Therefore, the relationship between HCV and RCC was investigated. The study included 100 patients with RCC; 32 with HCV infection, and 68 without HCV infection. Expressions of viral proteins (NS3 and NS5A) were tested using an immune electron-microscope (IEM) and immunohistochemistry (IHC). IHC and quantitative real time-PCR investigated the presentation of human proteins TP53 and p21 genes. Transmission electron (TEM) detected viral-like particles in infected RCC tissues. The gene and protein expression of P53 was higher in HCV positive versus HCV negative patients and p21 was lower in HCV positive versus HCV negative in both tumor and normal tissue samples. Viral like particles were observed by TEM in the infected tumor and normal portion of the RCC tissues and the plasma samples. The IEM showed the depositions of NS3 and NS5A in infected renal tissues, while in noninfected samples, were not observed. The study hypothesizes that a correlation between HCV and RCC could exist through successfully detecting HCV-like particles, HCV proteins, and (p53 and p21) in RCC-infected patients.

Exploring the antibiofilm and toxicity of tin oxide nanoparticles: Insights from in vitro and in vivo investigations.

BACKGROUND INFORMATION: The advancement of biological-mediated nanoscience towards higher levels and novel benchmarks is readily apparent, owing to the use of non-toxic synthesis processes and the incorporation of various additional benefits. This study aimed to synthesize stable tin oxide nanoparticles (SnO2-NPs) using S. rhizophila as a mediator. METHODS: The nanoparticles that were created by biosynthesis was examined using several analytical techniques, including Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), UV-visible (UV-vis) spectroscopy, and energy dispersive X-ray spectroscopy (EDS). RESULTS: The results obtained from the characterization techniques suggest that S. rhizophila effectively catalyzed the reduction of SnCl2 to SnO2-NPs duration of 90 min at ambient temperature with the ƛmax of 328 nm. The size of the nano crystallite formations was measured to be 23 nm. The present study investigates nanoscale applications' antibacterial efficacy against four bacterial strains, including Klebsiella Sp, Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli. The observed zone of inhibition for the nanoparticles (NPs) varied from 10 to 25 mm. The research findings demonstrate that the nanoparticles (NPs) are effective as antibacterial, phytotoxic, and cytotoxic agents.

Dual-channel neural network for instance segmentation of synapse.

Detection and segmentation of neural synapses in electron microscopy images are the committed steps for analyzing neural ultrastructure. To date, manual annotation of the structure in synapses has been the primary method, which is time-consuming and restricts the throughput of data acquisition. Recent studies have utilized a series of deformations based on a segmentation model for the detection and segmentation of transmission electron microscope images. However, the analysis of synaptic segmentation and statistics still lacks sufficient automation and high-throughput. Therefore, we developed a dual-channel neural network instance segmentation model with weighted top-down and multi-scale bottom-up schemes, which aid in accurately detecting and segmenting synaptic vesicles and their active zones within presynaptic membranes in complex environments. In addition, we proposed a masked self-supervised pre-training model based on the latest convolutional architecture to improve performance in downstream segmentation tasks. By comparing our model to other state-of-the-art methods, we determined its viability and accuracy. The applicability of our model is thoroughly demonstrated by distinct application scenarios for neurobiological research. These findings indicate that the dual-channel neural network could facilitate the analysis of synaptic structures for the advancement of biomedical research and electron microscope reconstruction techniques.

Resolving the In Situ Three-Dimensional Structure of Fly Mechanosensory Organelles Using Serial Section Electron Tomography.

Mechanosensory organelles (MOs) are specialized subcellular entities where force-sensitive channels and supporting structures (e.g., microtubule cytoskeleton) are organized in an orderly manner. The delicate structure of MOs needs to be resolved to understand the mechanisms by which they detect forces and how they are formed. Here, we describe a protocol that allows obtaining detailed information about the nanoscopic ultrastructure of fly MOs by using serial section electron tomography (SS-ET). To preserve fine structural details, the tissues are cryo-immobilized using a high-pressure freezer followed by freeze-substitution at low temperature and embedding in resin at room temperature. Then, sample sections are prepared and used to acquire the dual-axis tilt series images, which are further processed for tomographic reconstruction. Finally, tomograms of consecutive sections are combined into a single larger volume using microtubules as fiducial markers. Using this protocol, we managed to reconstruct the sensory organelles, which provide novel molecular insights as to how fly mechanosensory organelles work and are formed. Based on our experience, we think that, with minimal modifications, this protocol can be adapted to a wide range of applications using different cell and tissue samples. Key features • Resolving the high-resolution 3D ultrastructure of subcellular organelles using serial section electron tomography (SS-ET). • Compared with single-axis tilt series, dual-axis tilt series provides a much wider coverage of Fourier space, improving resolution and features in the reconstructed tomograms. • The use of high-pressure freezing and freeze-substitution maximally preserves the fine structural details.

A simple coordinate transformation method for quickly locating the features of interest in TEM samples.

We developed a simple coordinate transformation method for quickly locating features of interest (FOIs) of samples in transmission electron microscope (TEM). The method is well suited for conducting sample searches in aberration-corrected scanning/transmission electron microscopes (S/TEM), where the survey can be very time-consuming because of the limited field of view imposed by the highly excited objective lens after fine-tuning the aberration correctors. For implementation, a digital image of the sample and the TEM holder was captured using a simple stereo-optical microscope. Naturally presented geometric patterns on the holder were referenced to construct a projective transformation between the electron and optical coordinate systems. The test results demonstrated that the method was accurate and required no electron microscope or specimen holder modifications. Additionally, it eliminated the need to mount the sample onto specific patterned TEM grids or deposit markers, resulting in universal applications for most TEM samples, holders and electron microscopes for fast FOI identification. Furthermore, we implemented the method into a Gatan script for graphical-user-interface-based step-by-step instructions. Through online communication, the script enabled real-time navigation and tracking of the motion of samples in TEM on enlarged optical images with a panoramic view.

Controllable Pseudoelasticity in Metallic Nanocrystals by Grain Boundary Engineering.

Materials with pseudoelasticity can recover from large strains exceeding their elastic limits during unloading, making them promising damage-tolerant building blocks for advanced nanodevices. Nevertheless, a practical approach to realize controllable pseudoelastic behavior at nanoscale remains challenging. Here, we proposed a grain boundary (GB) engineering approach to endow metallic nanocrystals with a controllable pseudoelasticity. Both in situ nanomechanical testing and atomistic simulations demonstrate that such controllable pseudoelasticity is governed by the extension and contraction of an inherent stacking fault array at the GB. By precisely tuning GB misorientation and inclination, our simulation results reveal that metallic nanocrystals can exhibit tailored pseudoelastic performance across a broad spectrum of GBs in different face-centered cubic metals. These findings enrich our understanding of the intrinsic pseudoelasticity of GBs and provide a GB engineering approach toward metallic materials with reversible deformability.

Layered structure of sialoliths compared with tonsilloliths and antroliths.

Objectives: The aim of this study was to perform a comparative analysis of the ultrastructural and chemical composition of sialoliths, tonsilloliths, and antroliths and to describe their growth pattern. Materials and Methods: We obtained 19 specimens from 18 patients and classified the specimens into three groups: sialolith (A), tonsillolith (B), and antrolith (C). The peripheral, middle, and core regions of the specimens were examined in detail by histology, micro-computed tomography (micro- CT), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy, and transmission electron microscopy (TEM). Results: In the micro-CT, group A showed alternating radiodense and radiolucent layers, while group B had a homogeneous structure. Group C specimens revealed a compact homogeneous structure. Histopathologically, group A showed a laminated, teardrop-shaped, globular structure. Group B demonstrated degrees of immature calcification of organic and inorganic materials. In group C, the lesion was not encapsulated and showed a homogeneous lamellar bone structure. SEM revealed that group A showed distinct three layers: a peripheral multilayer zone, intermediate compact zone, and the central nidus area; groups B and C did not show these layers. The main elemental components of sialoliths were O, C, Ca, N, Cu, P, Zn, Si, Zr, F, Na, and Mg. In group B, a small amount of Fe was found in the peripheral region. Group C had a shorter component list: Ca, C, O, P, F, N, Si, Na, and Mg. TEM analysis of group A showed globular structures undergoing intra-vesicular calcification. In group B, bacteria were present in the middle layer. In the outer layer of the group C antrolith, an osteoblastic rimming was observed. Conclusion: Sialoliths had distinct three layers: a peripheral multilayer zone, an intermediate compact zone and the central nidus area, while the tonsillolith and antrolith specimens lacked distinct layers and a core.

Ultrastructural diversity in the shell of Artemia resting eggs.

The shell of Artemia resting egg, which is a delicate multilayered envelope surrounding the inside diapause embryo, plays an important role in the survival strategy of Artemia. To date, the ultrastructure of resting eggshell has been studied for only handful populations, and knowledge about the diversity of shell structure is still limited. In this paper, resting eggs from 13 Artemia populations were studied by transmission electron microscopy. Results show that the basic configuration of resting eggshell is quite conservative, but variations are not uncommon in the fine ultrastructure of each main layer of the shell (e.g., the shape and distribution of the radially oriented pores in the cortical layer; the size, number and arrangement of chambers in the alveolar layer; and the development state of outer cuticular membrane [OCM]). The ultrastructural variation of eggshell seems not to be linked with species and reproductive mode of Artemia. Resting eggs from very high habitats (4300+ m above sea level [a.s.l.]) on Qinghai-Tibet Plateau and certain tropical salterns have a hypoplastic OCM, which may be related to the adaptation to habitat conditions such as low oxygen concentration. RESEARCH HIGHLIGHTS: Comparative study on resting eggs from 13 Artemia populations reveals high diversity in the fine structure of eggshell. Resting eggs from very high (4300+ m a.s.l.) habitats commonly have a hypoplastic OCM.

Chemical and spectroscopic characterization of (Artemisinin/Querctin/ Zinc) novel mixed ligand complex with assessment of its potent high antiviral activity against SARS-CoV-2 and antioxidant capacity against toxicity induced by acrylamide in male rats.

A novel Artemisinin/Quercetin/Zinc (Art/Q/Zn) mixed ligand complex was synthesized, tested for its antiviral activity against coronavirus (SARS-CoV-2), and investigated for its effect against toxicity and oxidative stress induced by acrylamide (Acy), which develops upon cooking starchy foods at high temperatures. The synthesized complex was chemically characterized by performing elemental analysis, conductance measurements, FT-IR, UV, magnetic measurements, and XRD. The morphological surface of the complex Art/Q/Zn was investigated using scanning and transmission electron microscopy (SEM and TEM) and energy dispersive X-ray analysis (XRD). The in vitro antiviral activity of the complex Art/Q/Zn against SARS-CoV-2 and its in vivo activity against Acy-induced toxicity in hepatic and pulmonary tissues were analyzed. An experimental model was used to evaluate the beneficial effects of the novel Art/Q/Zn novel complex on lung and liver toxicities of Acy. Forty male rats were randomly divided into four groups: control, Acy (500 mg/Kg), Art/Q/Zn (30 mg/kg), and a combination of Acy and Art/Q/Zn. The complex was orally administered for 30 days. Hepatic function and inflammation marker (CRP), tumor necrosis factor, interleukin-6 (IL-6), antioxidant enzyme (CAT, SOD, and GPx), marker of oxidative stress (MDA), and blood pressure levels were investigated. Histological and ultrastructure alterations and caspase-3 variations (immunological marker) were also investigated. FT-IR spectra revealed that Zn (II) is able to chelate through C=O and C-OH (Ring II) which are the carbonyl oxygen atoms of the quercetin ligand and carbonyl oxygen atom C=O of the Art ligand, forming Art/Q/Zn complex with the chemical formula [Zn(Q)(Art)(Cl)(H2O)2]⋅3H2O. The novel complex exhibited a potent anti-SARS-CoV-2 activity even at a low concentration (IC50 = 10.14 µg/ml) and was not cytotoxic to the cellular host (CC50 = 208.5 µg/ml). Art/Q/Zn may inhibit the viral replication and binding to the angiotensin-converting enzyme-2 (ACE2) receptor and the main protease inhibitor (MPro), thereby inhibiting the activity of SARS-CoV-2 and this proved by the molecular dynamics simulation. It alleviated Acy hepatic and pulmonary toxicity by improving all biochemical markers. Therefore, it can be concluded that the novel formula Art/Q/Zn complex is an effective antioxidant agent against the oxidative stress series, and it has high inhibitory effect against SARS-CoV-2.

Transmission electron microscopic study of the surface layer of surgical resected disc specimens in human temporomandibular joint.

In this study, we investigated specific and characteristic findings of the surface layer of surgical resected disc specimens in human temporomandibular joint osteoarthritis cases by transmission electron microscopy (TEM).Specimens were surgically removed from the TMJ of 5 cases (4 female patients: 5 cases) clinically osteoarthritis. Following findings were observed by TEM. Images were photographed on a JEM1400-Flash Electron microscope (JEOL, Japan) equipped with an EM-14661FLASH high-sensitivity digital complementary metal-oxide-semiconductor camera.Following findings were observed by TEM. 1) The surface is covered with plump fibroblastic and histiocytoid cells. 2) Collagen fiber bundles and collagenous matrix are exposed onto the eroded disc surface. 3) Fibrinous dense material is observed on the eroded disc surface. 4) Bundles of collagen fibers are densely observed. 5) Collagen bundles are rich around capillary vessels. 6) Synovial surface cells reveal features of activated macrophages with vacuole formation. Especially, plump fibroblastic and histiocytoid cells, and activated macrophages with vacuole, which were significant findings of the surface layer. These findings might have a significant effect on the regulation of synovial fluid.

A Complicated Route from Disorder to Order in Antimony-Tellurium Binary Phase Change Materials.

The disorder-to-order (crystallization) process in phase-change materials determines the speed and storage polymorphism of phase-change memory devices. Only by clarifying the fine-structure variation can the devices be insightfully designed, and encode and store information. As essential phase-change parent materials, the crystallized Sb-Te binary system is generally considered to have the cationic/anionic site occupied by Sb/Te atoms. Here, direct atomic identification and simulation demonstrate that the ultrafast crystallization speed of Sb-Te materials is due to the random nature of lattice site occupation by different classes of atoms with the resulting octahedral motifs having high similarity to the amorphous state. It is further proved that after atomic ordering with disordered chemical occupation, chemical ordering takes place, which results in different storage states with different resistance values. These new insights into the complicated route from disorder to order will play an essential role in designing neuromorphic devices with varying polymorphisms.

Sensing Mechanism and Characterization of NO2 Gas Sensors Using Gold-Black NP-Decorated Ga2O3 Nanorod Sensing Membranes.

In this work, a vapor cooling condensation system was utilized to deposit various amounts of p-type gold-black nanoparticles (NPs) onto the surface of n-type gallium oxide (Ga2O3) nanorods forming p-n heterojunction-structured sensing membranes of nitrogen dioxide (NO2) gas sensors. The role and the sensing mechanism of the various gold-black NP-decorated Ga2O3 nanorods in NO2 gas sensors were investigated. The coverage and atomic percentage of the sensing membranes were observed using high-resolution transmission electron microscopy (HRTEM) measurements and energy-dispersive spectroscopy (EDS), respectively. For the NO2 gas sensor using the sensing membrane of 60 s-deposited gold-black NP-decorated Ga2O3 nanorods under a NO2 concentration of 10 ppm, the highest responsivity of 5221.1% was obtained. This result was attributed to the spillover effect and the formation of the p-n heterojunction, which increased more ionized-oxygen adsorption sites and promoted the reaction between NO2 gas and Ga2O3 nanorods. Furthermore, the NO2 gas sensor could detect the low NO2 concentration of 100 ppb and achieved a responsivity of 56.9%. The resulting NO2 gas sensor also exhibited excellent selectivity for detecting NO2 gas, with higher responsivity at a NO2 concentration of 10 ppm compared with that of the C2H5OH and NH3 concentrations of 100 ppm.

Buckling and fracture characterization of pristine bundles of vertically aligned carbon nanotubes using quantitativein situTEM axial compression.

This work investigates the mechanical deformation and fracture characteristics of pristine bundles of vertically aligned multi-walled carbon nanotubes (MWCNTs) subjected to axial compressionin situtransmission electron microscope (TEM). Accurate measurements of force-displacement data were collected simultaneously with real-time TEM videos of the deformation process. Two distinct regimes were observed in the force-displacement curve: (1) an initial elastic section with a linear slope, followed by (2) a transition to a force plateau at a critical buckling force. Morphological data revealed coordinated buckling of the pristine bundle, indicating strong van der Waals (VdW) forces between the nanotubes. The experimental setup measured an effective modulus of 83.9 GPa for an MWCNT bundle, which was in agreement with finite element analysis (FEA) simulations. FEA also highlighted the significant role of VdW forces in the bundle mechanical reactions. Furthermore, we identified nickel nanoparticles as key players in the fracture behavior of the bundles, acting as nucleation sites for defects. The direct mechanical measurements of MWCNT bundles provide valuable insights into their mechanical deformation and fracture behavior, while correlating it to the morphology of the bundle. Understanding these interactions at the bundle level is crucial for improving the reliability and durability of VACNTs-based components.

Evolution of Short-Range Order of Amorphous GeTe Upon Structural Relaxation Obtained by TEM Diffractometry and RMC Methods.

Glasses frequently reveal structural relaxation that leads to changes in their physical properties including enthalpy, specific volume, and resistivity. Analyzing the short-range order (SRO) obtained from electron diffraction by transmission electron microscopy (TEM) in combination with Reverse-Monte-Carlo (RMC) simulations is shown to provide information on the atomic arrangement. The technique elaborated here features several benefits including reliability, accessibility, and allows for obtaining detailed structural data quickly. This is demonstrated with a detailed view of the structural changes in the as-deposited amorphous phase change material (PCM) GeTe. The data show a significant increase in the average bond angle upon thermal treatment. At the same time the fraction of tetrahedrally coordinated Ge atoms decreases due to an increase in octahedrally distorted and pyramidal motifs. This finding provides further evidence for the atomic processes that govern structural relaxation in amorphous GeTe and other PCMs. A thorough literature review finally unveils possible origins of the large discrepancies reported on the structure of amorphous GeTe.

Pathological analysis of lesions in the exocrine pancreas of rats induced by Zinc Maltol.

The pancreas plays an important role in the homeostasis of zinc (Zn), a nutritionally essential metal. In several previous studies, Zn ions induced inflammatory changes in the exocrine pancreas; however, little is known about Zn complexes. In this study, we microscopically, immunohistochemically, and ultrastructurally examined pancreatic lesions in Sprague-Dawley (SD) rats induced by a 4-week repeated oral dose toxicity study of Zinc Maltol (ZM), a zinc (II) complex. ZM induces acinar atrophy and increases the number of duct-like structures. Immunohistochemistry revealed a decrease in the number of trypsin-positive cells, and an increase in the number of SOX9-positive cells. Interstitial fibrosis and macrophage infiltration also correlated with the degree of acinar atrophy. Electron microscopic evaluation revealed that the acinar cells that lost granules were surrounded by fibroblasts and collagen fibers. In conclusion, we provided a detailed description of ZM-induced pancreatic lesions in SD rats.

Morphological and ultrastructural studies of the internal reproductive systems of two deltocephaline leafhoppers, Nephotettixcincticeps and Deltocephalusvulgaris (Hemiptera, Cicadellidae, Deltocephalinae).

Insects have highly variable reproductive systems, reflecting a diversity of reproductive strategies and adaptations. Such variation has been widely used to classify and estimate phylogenetic relationships. Here, the morphology and ultrastructure of the internal reproductive systems of two leafhopper species are described and illustrated, using both light and transmission electron microscopy, and representing two tribes of Deltocephalinae: in Chiasmini, Nephotettixcincticeps (Uhler, 1896), and in Deltocephalini, Deltocephalusvulgaris (Dash & Viraktamath, 1998). Tables comparing the morphology of male and female internal reproductive structures of these studied species are provided and indicate that the main differences are in the relative shapes, sizes, and colors of these structures. The overall structure and organization, including details of the ultrastructure, of these two leafhopper species' male and female internal reproductive systems are very similar to those of previously studied leafhoppers. The main differences observed among species include the number of testicular follicles, the relative position of seminal vesicles and the degree of development of the accessory glands in the male, the number of ovaries, and the shape and color of the vagina and spermatheca in the female.

Investigation of the Efficacy of Lanthanoid Heavy Metal Acetates as Electron Staining Reagents for Biomembrane Vesicles.

Transmission electron microscopy (TEM) has revolutionized our understanding of protein structures by enabling atomic-resolution visualization without the need for crystallography, thanks to advancements in cryo-TEM and single particle analysis methods. However, conventional electron microscopy remains relevant for studying stained samples, as it allows the practical determination of optimal conditions through extensive experimentation. TEM also facilitates the examination of supramolecular complexes encompassing proteins, lipids, and nucleic acids. In this study, we investigated the applicability of lanthanoid reagents as electron-staining alternatives to uranyl acetate, which is globally regulated as a nuclear fuel material. We focus on a model biomembrane vesicle system, the chromatophores from the purple photosynthetic eubacterium Rhodospirillum rubrum, which integrate proteins and lipids. Through density distribution analysis of electron micrographs, we evaluated the efficacy of various lanthanoid acetates and found that triacetates of neodymium, samarium, and gadolinium exhibited similar staining effectiveness to uranyl acetate. Additionally, triacetates of praseodymium, erbium, and lutetium, followed by europium show promising results as secondary candidates. Our findings suggest that lanthanoid transition heavy metal acetates could serve as viable alternatives for electron staining in TEM, offering potential advantages over uranyl acetate.

Characterization of Anomalous Grains in FeCo Magnetic Alloys.

Heat-treated FeCo-based magnetic alloys were characterized using a suite of electron microscopy techniques to gain insight into their structural properties. Electron channeling contrast imaging (ECCI) in the scanning electron microscope (SEM) found unique grains towards the outer edge of a FeCo sample with nonuniform background contrast. High-magnification ECCI imaging of these nonuniform grains revealed a weblike network of defects that were not observed in standard uniform background contrast grains. High-resolution electron backscattered diffraction (HR-EBSD) confirmed these defect structures to be dislocation networks and additionally found subgrain boundaries within the nonuniform contrast grains. The defect content within these grains suggests that they are unrecrystallized grains, and ECCI can be used as a rapid method to quantify unrecrystallized grains. To demonstrate the insight that can be garnered via ECCI on these unique grains, the sample was imaged before and after micro indentation. This experiment showed that slip bands propagate throughout the material until interacting with the dislocation networks, suggesting that these specific defects provide a barrier to plastic deformation. Taken together, these results show how ECCI can be used to better understand failure mechanisms in alloys and provides further evidence that dislocation networks play a critical role in the brittle failure of FeCo alloys.