Is it really descemetocele? Morphology of extremely thin membrane that remained after severe corneal melting: a case report.

The aim of this study was to report transmission electron microscopic findings of a case with whole corneal descemetocele following infective corneal ulcer for the first time in literature. A 72-year-old male patient presented with infective corneal ulcer. After resolution of the infection, corneoscleral transplantation was performed. The excised very thin corneal membrane was processed for transmission electron microscopic examination. Transmission electron microscopic examination of the specimen revealed many layered structures that consisted of two different types of cells. The first type consisted of lighter staining polygonal cells, while the second consisted of elongated cells with relatively dense staining. All cells were connected with a large number of gap or adherens junctions with intercalation of the cell membranes of adjacent cells. A haphazard distribution of cytoplasmic microfilaments were also observed in all of the cell types. There was no evidence of the presence of endothelial cells throughout the specimen. There was also no evidence of Descemet membrane presence except for a small part adjacent to iris tissue that contained some melanosomes. Although we clinically diagnosed descemetocele, Descemet membrane was not present at the electron microscopic level, and thus, the expression "descemetocele" is inappropriate.

Obtaining 3D Atomic Reconstructions from Electron Microscopy Images Using a Bayesian Genetic Algorithm: Possibilities, Insights, and Limitations.

The Bayesian genetic algorithm (BGA) is a powerful tool to reconstruct the 3D structure of mono-atomic single-crystalline metallic nanoparticles imaged using annular dark field scanning transmission electron microscopy. The number of atoms in a projected atomic column in the image is used as input to obtain an accurate and atomically precise reconstruction of the nanoparticle, taking prior knowledge and the finite precision of atom counting into account. However, as the number of parameters required to describe a nanoparticle with atomic detail rises quickly with the size of the studied particle, the computational costs of the BGA rise to prohibitively expensive levels. In this study, we investigate these computational costs and propose methods and control parameters for efficient application of the algorithm to nanoparticles of at least up to 10 nm in size.

Colloidal platinum nanoparticles enhance resin-dentin bonding durability.

OBJECTIVES: This study aims to investigate the effect of colloidal platinum nanoparticles (CPN) on the durability of resin-dentin bonding performance with contemporary adhesives. METHODS: Sixty non-carious human maxillary premolars were subjected to microtensile bond strength (µTBS) testing and divided into two main groups: CPN-treated and untreated. Within each group, specimens were randomly allocated to Clearfil Megabond 2 (MB2), Scotchbond Universal Plus Adhesive with self-etch mode (SE-SUP), and etch-and-rinse mode (ER-SUP) subgroups (n = 10/group). CPN was applied to dentin in the MB2 and SE-SUP groups for 20 s, followed by rinsing before adhesive application. In the ER-SUP group, CPN was applied after etch-and-rinse. The µTBS was tested after 24 h, 6 months, and 1 year, and the fracture modes were observed using SEM. The µTBS data were analyzed using a two-way ANOVA and post-hoc Tukey HSD test (α = 0.05). An additional twelve premolars underwent TEM/STEM/EDX for ultra-morphological observations. RESULTS: The application of CPN significantly prevented a decline in the µTBS of both the MB2 and SE-SUP groups. No significant decrease was observed in the ER-SUP group, either with aging or CPN application. Ultra-morphological images revealed platinum nanoparticles attaching to the collagen fibrils of the hybrid layer regardless of aging. It was highlighted that the nanoparticles attached to the banded collagen in the aging groups were observed. SIGNIFICANCE: CPN exhibits the potential in enhancing the longevity of resin-dentin bonding in SE mode.

Precise Size Determination of Supported Catalyst Nanoparticles via Generative AI and Scanning Transmission Electron Microscopy.

Transmission electron microscopy (TEM) plays a crucial role in heterogeneous catalysis for assessing the size distribution of supported metal nanoparticles. Typically, nanoparticle size is quantified by measuring the diameter under the assumption of spherical geometry, a simplification that limits the precision needed for advancing synthesis-structure-performance relationships. Currently, there is a lack of techniques that can reliably extract more meaningful information from atomically resolved TEM images, like nuclearity or geometry. Here, cycle-consistent generative adversarial networks (CycleGANs) are explored to bridge experimental and simulated images, directly linking experimental observations with information from their underlying atomic structure. Using the versatile Pt/CeO2 (Pt particles centered ≈2 nm) catalyst synthesized by impregnation, large datasets of experimental scanning transmission electron micrographs and physical image simulations are created to train a CycleGAN. A subsequent size-estimation network is developed to determine the nuclearity of imaged nanoparticles, providing plausible estimates for ≈70% of experimentally observed particles. This automatic approach enables precise size determination of supported nanoparticle-based catalysts overcoming crystal orientation limitations of conventional techniques, promising high accuracy with sufficient training data. Tools like this are envisioned to be of great use in designing and characterizing catalytic materials with improved atomic precision.

Size-Dependent Core-Shell Fine Structures and Oxygen Evolution Activity of Electrochemical IrOx Nanoparticles Revealed by Cryogenic Electron Microscopy.

Electrochemically oxidized amorphous iridium oxides (IrOx) offer significantly improved electrocatalytic activities on the oxygen evolution reaction (OER) compared to crystalline IrO2, yet the origin of their decent activity and their size-dependent properties have not been fully understood. An important argument is the formation of deprotonated oxygen species not only at the topmost surface but also at the near surface, which creates an electrophilic character that activates the OER electrocatalysis. However, high spatial resolution identification of the electrophilic oxygen species remains unachieved. We address this hitherto-unresolved problem on size-selected electrochemical IrOx nanoparticles (NPs) by using cryogenic scanning transmission electron microscopy combined with electron energy loss spectroscopy, which enables simultaneous atomic detection of the near surface compositional and electronic structures with minimal damage that are further correlated with their size-dependent OER activities. Depending on the particle size, the electrochemical IrOx NPs showed distinctly different core-shell fine structures ranging from amorphous and hydrous IrOxHy NPs to a "metallic Ir core/sub-stoichiometric IrOx interlayer/amorphous IrOxHy shell" NP structure. Moreover, the formation of deprotonated, electrophilic oxygen is directly identified at the substoichiometric IrOx interface layer. These features account for a previously unestablished particle size effect of the electrochemical IrOx NPs, showing increasing water oxidation reactivity with an increasing nanoparticle size. Our results provide important insights into how subsurface oxygen chemistry controls the surface reactivity in the nanoscale Ir-based OER electrocatalysts.

Low-dose electron microscopy imaging for beam-sensitive metal-organic frameworks.

Metal-organic frameworks (MOFs) have garnered significant attention in recent years owing to their exceptional properties. Understanding the intricate relationship between the structure of a material and its properties is crucial for guiding the synthesis and application of these materials. (Scanning) Transmission electron microscopy (S)TEM imaging stands out as a powerful tool for structural characterization at the nanoscale, capable of detailing both periodic and aperiodic local structures. However, the high electron-beam sensitivity of MOFs presents substantial challenges in their structural characterization using (S)TEM. This paper summarizes the latest advancements in low-dose high-resolution (S)TEM imaging technology and its application in MOF material characterization. It covers aspects such as framework structure, defects, and surface and interface analysis, along with the distribution of guest molecules within MOFs. This review also discusses emerging technologies like electron ptychography and outlines several prospective research directions in this field.

Picometer-Level In Situ Manipulation of Ferroelectric Polarization in Van der Waals layered InSe.

Ferroelectric 2D van der Waals (vdW) layered materials are attracting increasing attention due to their potential applications in next-generation nanoelectronics and in-memory computing with polarization-dependent functionalities. Despite the critical role of polarization in governing ferroelectricity behaviors, its origin and relation with local structures in 2D vdW layered materials have not been fully elucidated so far. Here, intralayer sliding of approximately six degrees within each quadruple-layer of the prototype 2D vdW ferroelectrics InSe is directly observed and manipulated using sub-angstrom resolution imaging and in situ biasing in an aberration-corrected scanning transmission electron microscope. The in situ electric manipulation further indicates that the reversal of intralayer sliding can be achieved by altering the electric field direction. Density functional theory calculations reveal that the reversible picometer-level intralayer sliding is responsible for switchable out-of-plane polarization. The observation and manipulation of intralayer sliding demonstrate the structural origin of ferroelectricity in InSe and establish a dynamic structural variation model for future investigations on more 2D ferroelectric materials.

Unraveling Interdiffusion Phenomena and the Role of Nanoscale Diffusion Barriers in the Copper-Gold System.

Diffusion is one of the most fundamental concepts in materials science, playing a pivotal role in materials synthesis, forming, and degradation. Of particular importance is solid state interdiffusion of metals which defines the usable parameter space for material combinations in the form of alloys. This parameter space can be explored on the macroscopic scale by using diffusion couples. However, this method reaches its limit when going to low temperatures, small scales, and when testing ultrathin diffusion barriers. Therefore, this work transfers the principle of the diffusion couples to small scales by using core-shell nanowires and in situ heating. This allows us to delve into the interdiffusion dynamics of copper and gold, revealing the interplay between diffusion and the disorder-order phase transition. Our in situ TEM experiments in combination with chemical mapping reveal the interdiffusion coefficients of Cu and Au at low temperatures and highlight the impact of ordering processes on the diffusion behavior. The formation of ordered domains within the solid-solution is examined using high-resolution imaging and nanodiffraction including strain mapping. In addition, we examine the effectiveness of ultrathin Al2O3 barrier layers to control interdiffusion of the diffusion couple. Our findings indicate that a 5 nm thick layer serves as an efficient diffusion barrier. This research provides valuable insights into the interdiffusion behavior of Cu and Au on the nanoscale, offering potential applications in the development of miniaturized integrated circuits and nanodevices.

Resolution Improvement of Differential Phase-Contrast Microscopy via Tilt-Series Acquisition for Environmental Cell Application.

A simple method that improves the resolution of the phase measurement of differential phase-contrast (DPC) scanning transmission electron microscopy for closed-type environmental cell applications was developed and tested using a model sample simulating environmental cell observations. Because the top and bottom membranes of an environmental cell are typically far apart, the images from these membranes are shifted widely by tilt-series acquisition, and averaging the images after alignment can effectively eliminate undesired signals from the membranes while improving the signal from the object of interest. It was demonstrated that a phase precision of 2π/100 rad is well achievable using the proposed method for the sample in an environmental cell.

Unlocking Unexpected Charge Transfer Pathways in Interconnected Nanostructures.

Accurate control of charge transfer pathways is critical to unlocking the full potential of charge transfer proteins (CTPs) and exploring their diverse applications. We show that the intentional manipulation of junctions in Al nanocrosses on graphene induces asymmetry, unlocking unexpected charge transfer pathways and facilitating the generation of coupled resonators. The junction asymmetry, which is induced by nanotrench formation facilitated by focused electron beam irradiation, provides a versatile means to achieve precise and controlled interconnect manipulation. We find that tuning the nanotrench dimensions in nanocrosses allows for the tailored modulation of the charge transfer speed and the energies of CTPs. Furthermore, CTPs excited in our experimental nanocrosses, featuring nanotrenches, exhibit weak coupling. This crucial insight underscores the importance of controlled trench formation in unlocking various functionalities of CTPs for use in sensing, catalysis, and energy conversion applications. The controlled manipulation of interconnects in Al nanocrosses thus emerges as a promising avenue for advancing the device performance in these fields.

Nanoscale View of Alignment and Domain Growth in a Hexagonal Columnar Liquid Crystal.

Highly ordered liquid crystalline (LC) phases have important potential for organic electronics. We studied the molecular alignment and domain structure in a columnar LC thin film with nanometer resolution during in situ heating using four-dimensional scanning transmission electron microscopy (4D STEM). The initial disordered vapor-deposited LC glass thin film rapidly ordered at its glass transition temperature into a hexagonal columnar phase with small (<10 nm), well-aligned, planar domains (columns oriented parallel to the surface). Upon further heating, the domains coarsen via bulk diffusion, then the film crystallizes, then finally transforms back to an LC phase at an even higher temperature. The LC phase at high temperature shows straight columns of molecules, which we attribute to structure inherited from the intermediate crystalline phase. Nanoscale 4D STEM offers direct insight into the mechanisms of domain reorganization, and intermediate crystallization is a potential approach to manipulate orientational order and texture at the nano- to mesoscale in LC thin films.

Isolation of Orientia tsutsugamushi in Culture from Vellore, South India.

Background: Orientia tsutsugamushi, causative agent of scrub typhus is an obligate intracellular parasite. We present information on isolation of this pathogen at a tertiary care centre in Vellore, Southern India. Materials & Methods: PBMCs (peripheral blood mononuclear cells) collected from suspected scrub typhus patients were inoculated into Vero and L929 cell lines and incubated at 37°C with 5% CO2 for 30 days. They were examined for presence of Orientia tsutsugamushi on 10, 15, 20 days post-inoculation and everyday thereafter for a maximum of 30 days post inoculation. The scrapings were subjected to Giemsa staining, IFA, 47kDa qPCR and transmission electron microscopy (TEM). The isolates were passaged 3-4 times to ensure viability and then stored in DMEM with 10% FBS (-80). Genotyping of the isolates was performed by amplifying a 650 bp segment of the TSA 56 (type specific antigen 56) gene. Results: Amongst the 50 samples inoculated, three were culture positive as confirmed by 47 kDa qPCR on 24th day post inoculation. This was further confirmed by Giemsa, IFA staining and TEM. The 650bp amplicons showed 99.5 to 100% homology with Orientia tsutsugamushi MW604716, MH003839, MW604718, MW604717, MH922787 and MH003838 strains. Phylogenetic analysis revealed that 2 isolates belong to TA763 genotype and one belongs to Gilliam genotype. Conclusion: Orientia tsutsugamushi has been isolated for the first time at Vellore, South India from PBMCs. Complete genomic analysis will give more information.

Effect of silver nanoparticles on donkey sperm parameters and ultrastructure.

The aim of this study was to determine the effect of silver nanoparticles (AgNPs) on donkey sperm parameters and ultrastructure. AgNPs were synthesized, purified and resuspended in the extender. Nine frozen-thawed donkey sperm samples were exposed to different concentrations of AgNPs (0, 1.25, 2.5, 5, 12.5, 25 and 50 µg/mL). Sperm parameters: total (TMOT, %) and progressive (PMOT, %) sperm motility, plasma (LIVE, %) and acrosomal membrane integrity (AIS, %), and sperm morphology (MORF, %) were evaluated immediately after AgNPs exposure (T0) and after 2 h of incubation (T2). The interaction beween AgNPs and spermatozoa was visualized by transmission electron microscopy (TEM). At T0, sperm motility and AIS were reduced (p < .05) when using concentrations ≥50 and ≥25 µg/mL, respectively. At T2, sperm motility and LIVE were significantly decreased (p < .05) in concentrations ≥25 and ≥50 µg/mL, respectively. TEM analysis revealed nanoparticle adhesion to the acrosomal region of the plasma membrane. In conclusion, AgNPs at concentrations ≥25 µg/mL impair motility, acrosome and plasma membrane integrity of donkey sperm, which may be mediated by adhesion to the acrosomal region of the sperm surface membrane.

Cu-Atom Locations in Rocksalt SnTe Thermoelectric Alloy.

The development of functional thermoelectric materials requires direct evidence of dopants' locations to rationally design the electronic and phononic structure of the host matrix. In this study, Cs-corrected scanning transmission electron microscopy and energy dispersive X-ray spectroscopy is employed at the atomic scale to identify Cu atoms' locations in a Cu-doped SnTe thermoelectric alloy. It is revealed that Cu atoms in the rocksalt SnTe form solid solutions at both Sn and Te sites, contrary to their electronegativity order and the intentional Cu doping at Sn sites. Cu atoms are also located at the tetrahedral and crowdion sites of the face-centred cubic structure, with varying degrees of correlations. Such high flexibility of Cu atoms in the rocksalt SnTe offers diverse phonon-scattering mechanisms conducive to the ultra-low lattice thermal conductivity of singly Cu-doped SnTe. This study offers atomic-scale insights for achieving more precise dopant engineering, leading to the accelerated development of functional thermoelectric materials.

Graphene Bilayer as a Template for Manufacturing Novel Encapsulated 2D Materials.

Bilayer graphene (BLG) has recently been used as a tool to stabilize encapsulated single sheets of various layered materials and tune their properties. It was also discovered that the protecting action of graphene sheets makes it possible to synthesize completely new two-dimensional materials (2DMs) inside the BLG by intercalating various atoms and molecules. In comparison to the bulk graphite, BLG allows for easier intercalation and a much larger increase in the interlayer separation of the sheets. Moreover, it enables studying the atomic structure of the intercalated 2DM by using high-resolution transmission electron microscopy. In this review, we summarize the recent progress in this area, with a special focus on new materials created inside BLG. We compare the experimental findings with the theoretical predictions, pay special attention to the discrepancies, and outline the challenges in the field. Finally, we discuss unique opportunities offered by intercalation into 2DMs beyond graphene and their heterostructures.

Enhancing Semantic Segmentation in High-Resolution TEM Images: A Comparative Study of Batch Normalization and Instance Normalization.

Integrating deep learning into image analysis for transmission electron microscopy (TEM) holds significant promise for advancing materials science and nanotechnology. Deep learning is able to enhance image quality, to automate feature detection, and to accelerate data analysis, addressing the complex nature of TEM datasets. This capability is crucial for precise and efficient characterization of details on the nano-and microscale, e.g., facilitating more accurate and high-throughput analysis of nanoparticle structures. This study investigates the influence of batch normalization (BN) and instance normalization (IN) on the performance of deep learning models for semantic segmentation of high-resolution TEM images. Using U-Net and ResNet architectures, we trained models on two different datasets. Our results demonstrate that IN consistently outperforms BN, yielding higher Dice scores and Intersection over Union metrics. These findings underscore the necessity of selecting appropriate normalization methods to maximize the performance of deep learning models applied to TEM images.

Control of Grain Boundary Formation in Atomically Resolved Nanocrystalline Carbon Monolayers: Dependence on Electron Energy.

In this study, we explore the dynamics of grain boundaries in nanocrystalline carbon monolayers, focusing on their variation with electron beam energy and electron dose rate in a spherical and chromatic aberration-corrected transmission electron microscope. We demonstrate that a clean surface, a high-dose rate, and a 60 keV electron beam are essential for precise local control over the dynamics of grain boundaries. The structure of these linear defects has been evaluated using neural network-generated polygon mapping.

Cost-Effective Thermomechanical Processing of Nanostructured Ferritic Alloys: Microstructure and Mechanical Properties Investigation.

Nanostructured ferritic alloys (NFAs), such as oxide-dispersion strengthened (ODS) alloys, play a vital role in advanced fission and fusion reactors, offering superior properties when incorporating nanoparticles under irradiation. Despite their importance, the high cost of mass-producing NFAs through mechanical milling presents a challenge. This study delves into the microstructure-mechanical property correlations of three NFAs produced using a novel, cost-effective approach combining severe plastic deformation (SPD) with the continuous thermomechanical processing (CTMP) method. Analysis using scanning electron microscopy (SEM)-electron backscatter diffraction (EBSD) revealed nano-grain structures and phases, while scanning transmission electron microscopy (STEM)-energy dispersive X-ray spectroscopy (EDS) quantified the size and density of Ti-N, Y-O, and Cr-O fine particles. Atom probe tomography (APT) further confirmed the absence of finer Y-O particles and characterized the chemical composition of the particles, suggesting possible nitride dispersion strengthening. Correlation of microstructure and mechanical testing results revealed that CTMP alloys, despite having lower nanoparticle densities, exhibit strength and ductility comparable to mechanically milled ODS alloys, likely due to their fine grain structure. However, higher nanoparticle densities may be necessary to prevent cavity swelling under high-temperature irradiation and helium gas production. Further enhancements in uniform nanoparticle distribution and increased sink strength are recommended to mitigate cavity swelling, advancing their suitability for nuclear applications.

Glandular cell products in adult cestode: A new tale of tapeworm interaction with fish innate immune response.

The caryophyllidean tapeworm Caryophyllaeus brachycollis (Janiszewska, 1953) is indigenous to the Lake Blidinje in the west-central part of Bosnia-Herzegovina where it infects chub Squalius tenellus (Heckel, 1843). Of 22 chubs examined, 45% were infected with C. brachycollis and a total of 912 specimens of this worm were counted. Histopathological and ultrastructural investigations were conducted on interface region between chub intestine and cestode scolex. Different sizes of lipid droplets in cestode tegument, in interface region and in chub enterocytes were observed. C. brachycollis lacks any specialized attachment organs and with an expanded, flattened scolex goes deep in mucosal folds and firmly attaches to them. In the epithelium of fish intestine, near the site of worm attachment, a high number of mucous cells and several rodlet cells were noticed. Indeed, within the intestinal tunica propria-submucosa, beneath the site of scolex attachment, numerous neutrophils and mast cells were encountered. Transmission electron microscopy of the apical part of the scolex of C. brachycollis showed the occurrence of a multicellular, syncytial glandular complex, the scolex produced membrane-bound secretory granules and their fibrillar contents discharged by merocrine and apocrine secretion onto the host-parasite interface. Our results are among the first to provide evidence on the sophisticated relationship between fish intestine and amorphous-undefinable substance produced by scolex glandular complex.

Dynamic Deformation in Nuclear Graphite and Underlying Mechanisms.

Time-dependent deformation in nuclear graphite is influenced by the creation and migration of radiation-induced defects in the reactor environment. This study investigates the role of pre-existing defects such as point defect clusters and Mrozowski cracks in nuclear graphite IG-110. Separate specimens were irradiated with a 2.8 MeV Au2+ beam with a fluence of 4.38 × 1014 cm-2 and an 8 MeV C2+ beam with a fluence of 1.24 × 1016 cm-2. Microscopic specimens were either mechanically loaded inside a transmission electron microscope (TEM) or subjected to ex situ indentation-based creep loading. In situ TEM tests showed significant plasticity in regions highly localized around the Mrozowski cracks, resembling slip or ripplocation bands. Slip bands were also seen near regions without pre-existing defects but at very high stresses. Ex situ self-ion irradiation embrittled the specimens and decreased the creep displacement and rate, while heavy ion irradiation resulted in the opposite behavior. We hypothesize that the large-sized gold ions (compared to the carbon atoms) induced interplanar swelling as well as cross-plane channels for increased defect mobility. These findings illustrate the role of pre-existing defects in the dynamic relaxation of stresses during irradiation and the need for more studies into the radiation environment's impact on the mechanical response of nuclear graphite.