Transmission Electron Microscopy Methods Research Articles

The superiority of the photocatalytic and antibacterial performance of mechanochemically synthesized CdS nanoparticles over solvothermal-prepared ones

In this work, we have developed a facile, dry, and environmentally friendly mechanochemical method for the synthesis of cadmium sulfide (m-CdS) nanoparticles in a planetary ball mill using non-toxic precursors. Thiourea was for the first time used as a precursor of sulfide ion in room temperature solid state ball milling synthesis. For comparison of the mechanochemical approach with others, cadmium sulfide nanoparticles were also prepared using the solvothermal method (s-CdS). The crystal structure of cadmium sulfide nanoparticles was studied by XRD, the qualitative chemical band properties were examined by Raman scattering and x-ray photoelectron spectroscopy analysis, and particle morphology and microstructure were investigated by scanning electron microscopy and transmission electron microscopy methods. The sizes of the m-CdS nanoparticles had 5–6 nm in diameter, which is 10 times smaller than the diameter of s-CdS nanorods. The photocatalytic activities of as-prepared cadmium sulfide nanoparticles on the dye degradation and hydrogen production by water splitting were evaluated and the antibacterial activities were also tested. The photocatalytic activity of m-CdS was superior to that of s-CdS in the degradation of Orange II under visible light irradiation. Better results for m-CdS were also evidenced in photocatalytic experiments on hydrogen generation. The maximum rate of hydrogen evolution for m-CdS was 191.9 µmolg−1h−1 at the 120th min,while this indicator for s-CdS was only 58.0 µmolg−1h−1 at the same irradiation time.The better effect of m-CdS was evidenced also in an antibacterial study (namely against gram-positive bacteria).

Preparing of N-P dual-doped auricularia auricula carbon host by yeast fermentation and its application in Li-S batteries

A nitrogen-phosphorus dual-doped auricularia auricula carbon (NP-AC) matrix was prepared through hydrothermal treatment using natural auricularia auricula as raw material and medium solution. Yeast was introduced and cultured by the auricularia auricula liquid medium. The NP-AC/S composite was prepared through further freeze-drying, carbonization, activation and compounding with sulfur procedures. The composition and microstructure of samples was characterized by X-ray diffraction (XRD), raman and X-ray photoelectron spectroscopy (XPS). The morphology was observed under scanning electron microscope (SEM) and transmission electron microscopy (TEM) method. The specific surface area was analyzed using nitrogen adsorption/desorption test. The weight ratio of sulfur was measured through thermogravimetric (TG) analysis. The original structure and composition of biomass were improved by yeast fermentation and the auricularia auricula block structure with low porosity was modified. The electrochemical performance of NP-AC/S composite in Li-S batteries was systematically explored. After 100 cycles at 0.2C current density, there was still nearly 1000 mAh g−1 reversible capacity, holding a capacity retention rate of more than 84 %. The cycle performance and rate performance were both far better than the ordinary activated carbon materials.

Revealing the Dynamic Lithiation Process of Copper Disulfide by in Situ TEM.

Transition metal oxides, fluorides, and sulfides are extensively studied as candidate electrode materials for lithium-ion batteries driven by the urgency of developing next-generation higher energy density lithium batteries. These conversion-type electrode materials often require nanosized active materials to enable a "smooth" lithiation and de-lithiation process during charge/discharge cycles, determined by their size, structure, and phase. Herein, the structural and chemical changes of Copper Disulfide (CuS2) hollow nanoparticles during the lithiation process through an in situ transmission electron microscopy (TEM) method are investigated. The study finds the hollow structure of CuS2 facilitates the quick formation of fluidic Li2S "drops," accompanied by a de-sulfurization to the Cu7S4 phase. Meanwhile, the metallic Cu phase emerges as fine nanoparticles and grows into nano-strips, which are embedded in the Li2S/Cu7S4 matrix. These complex nanostructured phases and their spatial distribution can lead to a low de-lithiation barrier, enabling fast reaction kinetics.

Comparative study of ion milling techniques for the preparation of ceramic fibers in transmission electron microscopy

Sample preparation, essential to any analysis, remains an under-documented step. Preparation methods for transmission electron microscopy (TEM) are complex and lead to artifacts that need to be identified to avoid wrong conclusions about the sample's microstructure. Ion milling techniques are increasingly becoming the reference techniques to prepare thin foils. The possibilities of different ion beam processes for milling samples will be shown and compared, using ceramic carbon/carbide fibers, a material applied in many industrial applications, as a test specimen. This overview of ion milling preparation techniques will enable us to identify the advantages, disadvantages and parameters in order to best prepare these thin samples. It will be carried out for the community, and will also highlight a new preparation method for ceramic fibers to minimize the artefacts inherent in these techniques.

AdipoRon ameliorates the progression of heart failure with preserved ejection fraction via mitigating lipid accumulation and fibrosis

IntroductionObesity and imbalance in lipid homeostasis contribute greatly to heart failure with preserved ejection fraction (HFpEF), the dominant form of heart failure. Few effective therapies exist to control metabolic alterations and lipid homeostasis. ObjectivesWe aimed to investigate the cardioprotective roles of AdipoRon, the adiponectin receptor agonist, in regulating lipid accumulation in the two-hit HFpEF model. MethodsHFpEF mouse model was induced using 60 % high-fat diet plus L-NAME drinking water. Then, AdipoRon (50 mg/kg) or vehicle were administered by gavage to the two-hit HFpEF mouse model once daily for 4 weeks. Cardiac function was evaluated using echocardiography, and Postmortem analysis included RNA-sequencing, untargeted metabolomics, transmission electron microscopy and molecular biology methods. ResultsOur study presents the pioneering evidence that AdipoR was downregulated and impaired fatty acid oxidation in the myocardia of HFpEF mice, which was associated with lipid metabolism as indicated by untargeted metabolomics. AdipoRon, orally active synthetic adiponectin receptor agonist, could upregulate AdipoR1/2 (independently of adiponectin) and reduce lipid droplet accumulation, and alleviate fibrosis to restore HFpEF phenotypes. Finally, AdipoRon primarily exerted its effects through restoring the balance of myocardial fatty acid intake, transport, and oxidation via the downstream AMPKα or PPARα signaling pathways. The protective effects of AdipoRon in HFpEF mice were reversed by compound C and GW6471, inhibitors of AMPKα and PPARα, respectively. ConclusionsAdipoRon ameliorated the HFpEF phenotype by promoting myocardial fatty acid oxidation, decreasing fatty acid transport, and inhibiting fibrosis via the upregulation of AdipoR and the activation of AdipoR1/AMPKα and AdipoR2/PPARα-related downstream pathways. These findings underscore the therapeutic potential of AdipoRon in HFpEF. Importantly, all these parameters get restored in the context of continued mechanical and metabolic stressors associated with HFpEF.

Facile Synthesis of Stable Nanosized Silver Clusters in Plasma Discharge Under Ultrasonic Cavitation

This work is devoted to the study of plasma-chemical processes determined by the combination of the effect of thermally nonequilibrium, low-temperature plasma and intensive ultrasonic vibrations in the regime of intensive cavitation on liquid-phase media. This method for the realization of plasma-chemical transformations has been proven to be of significant interest and to have advantages for the creation of new nano-sized materials with special properties because it allows for varying the electrophysical and acoustic characteristics of the process when carrying out plasma-chemical reactions and fusion reactions. In this work, a novel facile technique for the synthesis of nanosized silver clusters in plasma discharge under ultrasonic cavitation is reported. Such a type of plasma involves the simultaneous effect of high-intensity cavitation and steady electric discharge in a liquid phase between electrodes of desired material. As a result, stable tiny silver nanoclusters with around a 1-nm size and a relatively narrow particle size distribution were obtained using toluene as a liquid medium. The nanoclusters were characterized with dynamic light scattering, transmission electron microscopy, electron diffraction, and optical methods. The results confirmed the formation of nanoclusters with an absorbance peak around 300 nm and the absence of 400-nm peaks typical for silver nanoparticles. Fluorescence tests allowed for establishing the amorphous structure of the synthesized nanoclusters occupying the intermediate position between few-atom nanoclusters and nanoparticles. The nanoclusters obtained were proven to be stable for more than 3 months. The experiments also revealed the possibility of performing high-temperature plasma-chemical reactions that can be applied in fusion technology.

Digital twins and deep learning segmentation of defects in monolayer MX2 phases

Developing methods to understand and control defect formation in nanomaterials offers a promising route for materials discovery. Monolayer MX2 phases represent a particularly compelling case for defect engineering of nanomaterials due to the large variability in their physical properties as different defects are introduced into their structure. However, effective identification and quantification of defects remain a challenge even as high-throughput scanning transmission electron microscopy methods improve. This study highlights the benefits of employing first principles calculations to produce digital twins for training deep learning segmentation models for defect identification in monolayer MX2 phases. Around 600 defect structures were obtained using density functional theory calculations, with each monolayer MX2 structure being subjected to multislice simulations for the purpose of generating the digital twins. Several deep learning segmentation architectures were trained on this dataset, and their performances evaluated under a variety of conditions such as recognizing defects in the presence of unidentified impurities, beam damage, grain boundaries, and with reduced image quality from low electron doses. This digital twin approach allows benchmarking different deep learning architectures on a theory dataset, which enables the study of defect classification under a broad array of finely controlled conditions. It thus opens the door to resolving the underpinning physical reasons for model shortcomings and potentially chart paths forward for automated discovery of materials defect phases in experiments.

FBXL20 promotes synaptic impairment in depression disorder via degrading vesicle-associated proteins

BackgroundSynaptic plasticity changes in presynaptic terminals or postsynaptic membranes play a critical role in cognitive impairments and emotional disorders, but the underlying molecular mechanisms in depression remain largely unknown. MethodsThe regulation effects of F-box and leucine-rich repeat protein 20 (FBXL20), vesicular glutamate transporter 1 (VGLUT1) and vesicle-associated membrane protein 1 (VAMP1) on synaptic plasticity and depressive-like behaviors examined by proteomics analysis, viral stereotaxic injection, transmission electron microscope and biochemical methods. The glutamate release detected by fluorescent probe in cultured primary pyramidal neurons. ResultsWe found that chronic unpredictable mild stress (CUMS) induced significant synaptic deficits within hippocampus of depressed rats, accompanied with the decreased expression of VGLUT1 and VAMP1. Moreover, knockdown of VGLUT1 or VAMP1 in hippocampal pyramidal neurons resulted in abnormal glutamatergic neurotransmitter release. In addition, we found that the E3 ubiquitin ligase FBXL20 was increased within hippocampus, which may promote ubiquitination and degradation of VGLUT1 and VAMP1, and thus resulted in the reduction of glutamatergic neurotransmitter release, the disruptions of synaptic transmission and the induction of depression-like behaviors in rats. In contrast, shRNA knockdown of FBXL20 within the hippocampus of depressed rats significantly ameliorated synaptic damage and depression-like behaviors. LimitationOnly one type of depression model was used in the present study, while other animal models should be used in the future to confirm the underlying mechanisms reported here. ConclusionsThis study provides new insights that inhibiting FBXL20 pathway in depressed rats may be an effective strategy to rescue synaptic transmission and depression-like behaviors.

Visualisation and Evaluation Study of Some Patients Arteriosclerosis Microvessel with Coronary Artery Disease at Coronary Artery Bypass Grafting

Visualization of coronary microvessel at ischemic disease still remains actual. There is necessity for additional study of arteriosclerosis microvessels. The byoptates of auricular right atria of patients with coronary artery disease were taken at coronary artery bypass grafting. The epoxy slices were obtained from material treated by the method of transmission electron microscopy and fixed in epoxy resins with further staining by Azur II and viewed under light optical microscope. Analysis of the images obtained during this study identified different arteriosclerotic damages, with negative phenomena of convolute formation in the walls of arteriolas and formation of de Novo arteriolas from previous one, by intussusception type that could also lead to ischemia. Evaluation on 5-degree scale of pathological growth of vessels microcirculatory bed as well as provided 1-2 images could help during analyze of arteriosclerotic damages, identification of pathological process direction in each patient case. This will improve medical treatment of such patients in postsurgical period.

Surfactant modified coconut husk fiber as a green alternative sorbent for micro-solid phase extraction of triazole fungicides at trace level in environmental water, soybean milk, fruit juice and alcoholic beverage samples.

In this work, micro-solid phase extraction using surfactant modified biosorbent was investigated for trace level determination of triazole fungicides prior to their analysis by high performance liquid chromatography. Coconut husk fiber (CHF) was selected as an effective biosorbent in the extraction process. Fourier transform infrared spectrometry, scanning electron microscopy and transmission electron microscopy methods were used to characterize the modified biosorbent. Various factors affecting the extraction efficiency of the proposed method were studied including the amount of coconut husk fiber biosorbent (0.1 g), kind and concentration of surfactant as a modifier (sodium dodecyl sulfate, 10 mmol L-1), kind and volume of desorption solvent (methanol, 150 μL), and extraction period (including vortex adsorption time, centrifugation adsorption time, vortex desorption time and centrifugation adsorption time approximately 10 min). Under the selected conditions, the calibration plot was found to be linear in the range of 9-300 μg L-1 with a coefficient for determination of greater than 0.99. The limits of detection and limits of quantification for the studied triazole fungicides were 3.00 and 9.00 μg L-1, respectively. Finally, the proposed method was successfully applied to determine triazole fungicides in environmental water, soybean milk, fruit juice and alcoholic beverage samples with acceptable recoveries obtained in the range of 67.0% to 105.0%.

The role of high-resolution transmission electron microscopy and aberration corrected scanning transmission electron microscopy in unraveling the structure-property relationships of Pt-based fuel cells electrocatalysts.

Platinum-based fuel cell electrocatalysts are structured on a nano level in order to extend their active surface area and maximize the utilization of precious and scarce platinum. Their performance is dictated by the atomic arrangement of their surface layers atoms via structure-property relationships. Transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) are the preferred methods for characterizing these catalysts, due to their capacity to achieve local atomic-level resolutions. Size, morphology, strain and local composition are just some of the properties of Pt-based nanostructures that can be obtained by (S)TEM. Furthermore, advanced methods of (S)TEM are able to provide insights into the quasi-in situ, in situ or even operando stability of these nanostructures. In this review, we present state-of-the-art applications of (S)TEM in the investigation and interpretation of structure-activity and structure-stability relationships.

Change in structure and properties of high-speed steel deposited layer after tempering and electron beam processing

The structure and properties of multilayer coatings made of high-speed R18Yu steel formed on a substrate of 30KhGSA steel during multilayer plasma surfacing in a mixture of argon and nitrogen followed by high-temperature tempering and electron beam processing are studied using the methods of light, scanning and transmission electron microscopy. Plasma surfacing forms a layer, the main phases of which are α-Fe and carbides of М6С(Fe3W3C–Fe4W2C) composition, forming a frame mesh. After high-temperature tempering, particles of the carbide phase of globular or faceted shape are identified in the bulk of α-Fe with sizes from 10 to 38 nm. The microhardness of the deposited layer made of R18Yu steel increases after tempering and is 5.3 GPa. Electron beam processing is accompanied by a multiple (3 or more times) reduction in the wear parameter (increased wear resistance of the material); the microhardness of the modified layer does not change and is 5.3 GPa.

Photocatalysis oxidative desulfurization of dibenzothiophene in extremely deep liquid fuels on the Z-scheme catalyst ZnO-CuInS2-ZnS intelligently integrated with carbon quantum dots: performance, mechanism, and stability.

In this study, we improved the electrochemical and photocatalytic properties of the ZnO-CuInS2-ZnS (ZCZ) material by integrating with carbon quantum dots (CQD) with particle sizes from 2 to 5 nm. The integration of ZnO-CuInS2-ZnS with carbon quantum dots (ZnO-CuInS2-ZnS/CQD:ZCZ-CQD) enhanced the visible light absorption, significantly reduced the electron-hole recombination rate, and facilitated the electron transfer and separation processes as confirmed by UV-visible diffuse reflectance spectroscopy (UV-vis DRS), photoluminescence (PL), and electrochemical impedance spectroscopy (EIS). The successful integration of ZCZ with carbon quantum dots was confirmed using X-ray photoelectron spectroscopy (XPS), energy dispersive spectroscopy (EDS) and transmission electron microscopy (TEM) methods. The ZCZ/CQD photocatalyst removed up to 98.32% of DBT after 120 minutes of reaction, maintained over 90% durability after 10 cycles, and retained its structure without any changes. The ZCZ photocatalyst integrated with CQD enhances faster dibenzothiophene (DBT) removal by 4.46, 3.24, 2.53, and 1.72 times compared to ZnO, CuInS2, ZnS, and ZnO-CuInS2-ZnS, respectively. Factors influencing the oxidation process of DBT including the mass of the photocatalyst, initial DBT concentration, stability, and reaction kinetics were studied. Through active species trapping experiments, this study demonstrated that the formation of ˙O2 - and ˙OH radicals determines the reaction rate. The mechanism of photocatalysis on ZCZ-CQD materials and the intermediate products formed in the process of photocatalytic oxidative desulfurization of dibenzothiophene is proposed based on electrochemical measurements and GC-MS results.

Revealing microscopic dynamics: in situ liquid-phase TEM for live observations of soft materials and quantitative analysis via deep learning.

In various domains spanning materials synthesis, chemical catalysis, life sciences, and energy materials, in situ transmission electron microscopy (TEM) methods exert a profound influence. These methodologies enable the real-time observation and manipulation of gas-phase and liquid-phase reactions at the nanoscale, facilitating the exploration of pivotal reaction mechanisms. Fundamental research areas like crystal nucleation, growth, etching, and self-assembly have greatly benefited from these techniques. Additionally, their applications extend across diverse fields such as catalysis, batteries, bioimaging, and drug delivery kinetics. However, the intricate nature of 'soft matter' presents a challenge due to the unique molecular properties and dynamic behavior of these substances that remain insufficiently understood. Investigating soft matter within in situ liquid-phase TEM settings demands further exploration and advancement compared to other research domains. This research harnesses the potential of in situ liquid-phase TEM technology while integrating deep learning methodologies to comprehensively analyze the quantitative aspects of soft matter dynamics. This study centers on diverse phenomena, encompassing surfactant molecule nucleation, block copolymer behavior, confinement-driven self-assembly, and drying processes. Furthermore, deep learning techniques are employed to precisely analyze Ostwald ripening and digestive ripening dynamics. The outcomes of this study not only deepen the understanding of soft matter at its fundamental level but also serve as a pivotal foundation for developing innovative functional materials and cutting-edge devices.

LaFe1-xNixO3 perovskites in the catalytic reaction of high-temperature decomposition of nitrous oxide.

The catalytic performance of LaFe1-xNixO3 perovskites prepared by the Pechini method was studied in the high-temperature decomposition reaction of nitrous oxide at temperatures of 750-900 °C. The samples were characterized by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), transmission electron microscopy (TEM), adsorption (Sbet) and hydrogen thermo-programmed reduction (H2-TPR) methods. The prepared single-phase solids with specific surface areas of 4.7-8.4 m2 g-1 are porous particles (aggregates) of 10-100 μm composed of microblock crystallites. The surface of all samples is enriched with lanthanum. Increasing the Ni content increases the normalized per m2 catalytic activity of LaFe1-xNixO3 perovskites, their reducibility in hydrogen, the ratio of Fe + Ni/La and the content of low bond oxygen forms on the sample surface. The correlation of activity and low bond oxygen forms was revealed, which is consistent with the mechanism of reaction of nitrous oxide decomposition. For future industrial applications the LaFe0.4Ni0.6O3 perovskite is proposed due to its higher stability in the reaction medium as compared with samples with x > 0.6.

Membraneless ethanol fuel cell Pt-Sn-Re nano active catalyst on a mesoporous carbon support.

Herein, we report, for the first-time, mesoporous carbon-supported binary and ternary catalysts with different atomic ratios of Pt/MC (100), Pt-Sn/MC (50 : 50), Pt-Re/MC (50 : 50), Pt-Sn-Re/MC (80 : 10 : 10) and Pt-Sn-Re/MC (80 : 115 : 05) prepared using a co-impregnation reduction method as anode components for membraneless ethanol fuel cells (MLEFLs). Mechanistic and structural insights into binary Pt-Sn/MC, Pt-Re/MC and ternary Pt-Sn-Re/MC catalysts were obtained using scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and energy-dispersive X-ray spectroscopy (EDX) methods. In particular, chemical characterization via cyclic voltammetry, CO stripping voltammetry and chronoamperometry indicated that Pt-Sn-Re/MC (80 : 15 : 05) had better dynamics toward ethanol oxidation than Pt-Sn-Re/MC (80 : 10 : 10), Pt-Sn/MC (50 : 50) and Pt-Re/MC (50 : 50) catalysts. In terms of the single cell performance of the prepared catalysts, Pt-Sn-Re/MC (80 : 15 : 05) (31.5 mW cm-2) showed a higher power density and current density than Pt-Sn-Re/MC(80 : 10 : 10), Pt-Re/MC (50 : 50) and Pt-Sn/MC (50 : 50) at room temperature. The addition of Re into the binary Pt-Sn catalyst improved its electrical performance for ethanol oxidation in a membraneless ethanol fuel cell. As a result, the ternary-based Pt-Sn-Re/MC (80 : 15 : 05) catalyst demonstrated enhanced performance compared to monometallic and bimetallic catalysts in the ethanol oxidation reaction in a membraneless fuel cell.

Influence of shortwave laser therapy on morphometric parameters of sciatic nerve regeneration after experimental neurorrhaphy

The aim of this work was to reveal the effectiveness of delayed laser therapy for the regeneration of the sciatic nerve. Materials and methods. In 21 rats, transection and epineural microsurgical suture of the right sciatic nerve (SN) were performed. In 16 rats (experimental group) in the period from 1.0 to 2.5 months after surgery, a course of laser therapy (LT-1, n = 8) and a delayed (after 6 months) course of laser therapy (LT-2, n = 8) were used 18 sessions were performed in each subgroup. Laser therapy was not performed in 5 operated animals and they were included in the control group. The terms of withdrawal of rats from the experiment were 2.5 and 12.0 months after the operation. During the morphometry of semi-thin sections distal to the level of the sutures, the volumetric density of the constituent elements of the SN, the number density of myelinated nerve fibers (MNF), their diameters, the thickness of the axons and myelin sheath, the myelination index (index G) distribution of MNF by diameter and dependence of index G on the fiber caliber were measured. For double control, we used muscle morphometry data from intact rats (Intact). Muscle tissue was additionally examined by the method of transmission electron microscopy using a generally accepted technique with a REM-100 electron microscope (Selmi, Sumy, Ukraine). Results. In the control group, as compared to the intact group, the numerical density of muscle fibers was 21 % higher, but 38 % less than the numerical density of microvessels, therefore, the vascularization index was reduced by 1.7 times. The obtained results indicated laser compensation for denervation atrophy, which was confirmed by morphometric data. Conclusions. The results of the study indicate a stimulating prolonged effect of delayed laser therapy in regenerative axonogenesis.

Cryoconveyor protocols in correlation light and electron microscopy: From multilevel imaging to modeling of biophysical effects and “cryotheranostics”

This paper is a technical and methodological note, the purpose of which is to introduce into the practice of biological research methods of cryomicroscopy in a conveyor mode, starting from small magnifications and ending with the limits of magnification/resolution of scanning electron cryomicroscopy. The protocol described can be applied to the samples with low sample preparation complexity without ultratomy or the sample processing typical for transmission electron microscopy methods. According to this protocol samples are analyzed in a single microcuvette (chip) indexed by laboratory information management system and sequentially moved from the non-destructive low-resolution optical microscopy instruments (such as lensless cryomicroscopes) and optical super-resolution methods (some microinterferometers and MIMs with cryotables) to the CryoSEM/CryoESEM level (in programmable environments and atmospheres). Methods of correlation lensless cryomicroscopy and scanning microscopy (including those with the subsequent transition to microanalysis) were introduced; CryoCUVEM and CryoCIREM methods in the ultraviolet and infrared range, respectively; microinterferometry methods using a multi-beam reflected light interferometer (based on the MII-11 platform with several changes); the development of CryoCDICEM systems based on the optical path of an inverted metallographic microscope with a DIC attachment and a LED emitter was also initiated. The advantages of cryoconveyor analysis protocols are ensuring the sample safety in a single cuvette-chip and the possibility of establishing spatial colocalization between the data of optical and electron microscopy (including in the CLEM/CryoCLEM mode), as well as providing a comprehensive non- destructive sample analysis in the sequential study of the microscopic systems with the possibility of varying the subsequent stages of high-resolution microscopy, depending on the results obtained at the previous stages of lower resolution microscopic studies.

Observation and quantitative analysis of dislocations in steel using electron channeling contrast imaging method with precise control of electron beam incident direction.

Electron channeling contrast imaging (ECCI) was applied by precisely controlling the primary electron beam incident direction of the crystal plane in scanning electron microscope (SEM), and the dislocation contrast in steel materials was investigated in detail via SEM/ECCI. The dislocation contrast was observed near a channeling condition, where the incident electron beam direction of the crystal plane varied, and the backscattered electron intensity reached a local minimum. Comparing the dislocation contrasts in the visualized electron channeling contrast (ECC) images and transmission electron microscope (TEM) images, the positions of all dislocation lines were coincident. During the SEM/ECCI observation, the dislocation contrast varied depending on the incident electron beam direction of the crystal plane and accelerating voltages, and optimal conditions existed. When the diffraction condition g and the Burgers vector b of dislocation satisfied the condition g⸱b = 0, the screw dislocation contrast in the ECC image disappeared. An edge dislocation line was wider than a screw dislocation line. Thus, the SEM/ECCI method can be used for dislocation characterization and the strain field evaluation around dislocation, like the TEM method. The depth information of SEM/ECCI, where the channeling condition is strictly satisfied, can be obtained from dislocation contrast deeper than 5ξg, typically used for depth of SEM/ECCI.

Ultrastructure of neuron-glia interaction in the norm and experimental pathology

Long-term experience was gained in analyzing synapses and their glial surroundings in normal, natural, and experimental models of functional plasticity and brain pathology development. However, most studies in this area rely on electrophysiological techniques combined with fluorescent imaging. Notably, fine synapse structure and astrocytic processes cannot be resolved using light or some electron microscopy techniques. Studies on experimental brain pathology using volume electron microscopy methods [1] were previously restricted due to their high labor intensity. However, automated methods for sample preparation and analysis, using machine vision and artificial intelligence, significantly simplified this task. Using transmission electron microscopy methods and 3D reconstructions, this study examined the Str. radiatum CA1 hippocampal region of rat brains in a chronic lithium-pilocarpine model of epilepsy. The results indicate a decrease in synaptic number along with an increase in their size and a reduction in astrocytic isolation of the active zones. A decrease in glial ensheathment of enlarged active zones and facilitation of neurotransmitter diffusion to active synapses may have a multiplicative effect on epileptiform activity growth and excitotoxicity. The simplification of the astrocytes meshwork in the somatosensory cortex’s layer 2/3 is comparable to that of layer 1. This decrease in layer 1’s inhibitory action enables pyramidal neurons in layer 2/3 to potentially exhibit epileptiform activity. Thus, the superficial cortical layers’ structural-functional aspects can be used as a natural cellular model in epilepsy development studies. Reduction of Ca2+ events in astrocyte processes in lithium-pilocarpine induced epilepsy may result from the low buffer capacity of Ca2+ ions in the smooth endoplasmic reticulum (sER) and/or impaired Ca2+ wave transmission through the gap junctions between astrocytic processes. High resolution is necessary to analyze the gap junctions, and special methods are required for sER visualization in perisynaptic astrocytic processes. We developed original sER staining methods [2] to quantitatively evaluate gap junctions and sER cisternae within astrocytic meshworks in layers 1 and 2/3 of the somatosensory cortex. In layer 1, the area of gap junctions in relation to the volume of an astrocyte was twice as high as in layer 2/3. The proportion of sER volume differed between layer 1 and layer 2/3 tissues. Specifically, the total sER cisternae volume in layer 2/3 was twice as high as in layer 1, relative to the volume of astrocytic processes [3]. Additionally, a doubling of single astrocytic gap junction area concomitant with decreased calcium events was observed. The results suggest a normal balance between Ca2+ stores (sER) and gap junctions, whose disruption may contribute to the development of seizures.