Ion Beam Scanning Electron Microscopy Research Articles

Three-dimensional correlative microscopy of the Drosophila female reproductive tract reveals modes of communication in seminal receptacle sperm storage

In many taxa, females store sperm in specialized storage organs. Most insect sperm storage organs have a tubular structure, typically consisting of a central lumen surrounded by epithelial cells. These specialized tubules perform the essential tasks of transporting sperm through the female reproductive tract and supporting long-term sperm survival and function. Little is known about the way in which female sperm storage organs provide an environment conducive to sperm survival. We address this using a combined light microscopy, micro computed tomography (microCT), and Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) approach for high-resolution correlative three-dimensional imaging to advance our understanding of sperm-female interactions in Drosophila melanogaster. Using this multimodal approach, we were able to scan the lower female reproductive tract and distal portion of the seminal receptacle at low magnification, and to subsequently zoom in for further analysis on an ultrastructural level. Our findings highlight aspects of the way in which the seminal receptacle keeps sperm viable in the lumen, and set the stage for further studies. The methods developed are suitable not only for Drosophila but also for other organisms with soft, delicate tissues.

Quantitative characterization of organic and inorganic pores in shale based on deep learning

Organic matter (OM) maturity is closely related to organic pores in shales. Quantitative characterization of organic and inorganic pores in shale is crucial for rock-physics modeling and reservoir porosity and permeability evaluation. Focused ion beam-scanning electron microscopy (FIB-SEM) can capture high-precision three-dimensional (3D) images and directly describe the types, shapes, and spatial distribution of pores in shale gas reservoirs. However, due to the high scanning cost, wide 3D view field, and complex microstructure of FIB-SEM, more efficient segmentation for the FIB-SEM images is required. For this purpose, a multiphase segmentation workflow in conjunction with a U-net is developed to segment pores from the matrix and distinguish organic pores from inorganic pores simultaneously in the entire 3D image stack. The workflow is repeated for FIB-SEM data sets of 17 organic-rich shales with various characteristics. The analysis focuses on improving the efficiency and relevance of the workflow, that is, quantifying the minimum number of training slices while ensuring accuracy and further combining the fractal dimension (FD) and lacunarity to study a simple and objective method of selection. Meanwhile, the computational efficiency, accuracy, and robustness to noise of the 2D U-net model are discussed. The intersection over the union of automatic segmentation can amount to 80%–95% in all data sets with manual labels as ground truth. In addition, calculated by the FIB-SEM multiphase segmentation, the organic porosity is used to quantitatively evaluate the OM decomposition level. Deep-learning-based segmentation shows great potential for characterizing shale pore structures and quantifying OM maturity.

Focused ion beam-SEM 3D study of osteodentin in the teeth of the Atlantic wolfish Anarhichas lupus

The palette of mineralized tissues in fish is wide, and this is particularly apparent in fish dentin. While the teeth of all vertebrates except fish contain a single dentinal tissue type, called orthodentin, dentin in the teeth of fish can be one of several different tissue types. The most common dentin type in fish is orthodentin. Orthodentin is characterized by several key structural features that are fundamentally different from those of bone and from those of osteodentin. Osteodentin, the second-most common dentin type in fish (based on the tiny fraction of fish species out of ∼30,000 extant fish species in which tooth structure was so far studied), is found in most Selachians (sharks and rays) as well as in several teleost species, and is structurally different from orthodentin.Here we examine the hypothesis that osteodentin is similar to anosteocytic bone tissue in terms of its micro- and nano-structure. We use Focused Ion Beam-Scanning Electron Microscopy (FIB/SEM), as well as several other high-resolution imaging techniques, to characterize the 3D architecture of the three main components of osteodentin (denteons, inter-denteonal matrix, and the transition zone between them). We show that the matrix of osteodentin, although acellular, is extremely similar to mammalian osteonal bone matrix, both in general morphology and in the three-dimensional nano-arrangement of its mineralized collagen fibrils. We also document the presence of a complex network of nano-channels, similar to such networks recently described in bone. Finally, we document the presence of strings of hyper-mineralized small ‘pearls’ which surround the denteonal canals, and characterize their structure.

A Dynamic Permeability Model in Shale Matrix after Hydraulic Fracturing: Considering Mineral and Pore Size Distribution, Dynamic Gas Entrapment and Variation in Poromechanics

Traditional research on apparent permeability in shale reservoirs has mainly focussed on effects such as poromechanics and porosity-assisted adsorption layers. However, for a more realistic representation of field conditions, a comprehensive multi-scale and multi-flowing mechanism model, considering the fracturing process, has not been thoroughly explored. To address this research gap, this study introduces an innovative workflow for dynamic permeability assessment. Initially, an accurate description of the pore size distribution (PSD) within three major mineral types in shale is developed using focussed ion beam-scanning electron microscopy (FIB-SEM) and nuclear magnetic resonance (NMR) data. Subsequently, an apparent permeability model is established by combining the PSD data, leading to the derivation of dynamic permeability. Finally, the PSD-related dynamic permeability model is refined by incorporating the effects of imbibition resulting from the fracturing process preceding shale gas production. The developed dynamic permeability model varies with pore and fracture pressures in the shale reservoir. The fracturing process induces water blockage, water-film formation, and water-bridging phenomena in shale, requiring additional pressure inputs to counteract capillary effects in hydrophilic minerals in shale, But also increases the overall permeability from increasing permeability at larger scale pores. Unlike traditional reservoirs, the production process commences when the fracture is depleted to 1–2 MPa exceeds the pore pressure, facilitated by the high concentration of hydrophobic organic matter pores in shale, this phenomenon explains the gas production at the intial production stage. The reduction in adsorption-layer thickness resulting from fracturing impacts permeability on a nano-scale by diminishing surface diffusion and the corresponding slip flow of gas. this phenomenon increases viscous-flow permeability from enlarged flow spacing, but the increased viscous flow does not fully offset the reduction caused by adsorbed-gas diffusion and slip flow. In addition to the phenomena arising from various field conditions, PSD in shale emerges as a crucial factor in determining dynamic permeability. Furthermore, considering the same PSD in shale, under identical pore spacing, the shape factor of slit-like clay minerals significantly influences overall permeability characteristics, much more slit-shaped pores(higher shape factor) reduce the overall permeability. The dynamic permeability-assisted embedded discrete fracture model (EDFM) showed higher accuracy in predicting shale gas production compared to the original model.

UV-assisted nanoimprint lithography: the impact of the loading effect in silicon on nanoscale patterns of metalens.

This work studies the impact of the silicon (Si) loading effect induced by deep reactive ion etching (DRIE) of silicon master molds on the UV-nanoimprint lithography (NIL) patterning of nanofeatures. The silicon molds were patterned with metasurface features with widths varying from 270 to 60 nm. This effect was studied by focus ion beam scanning electron microscopy (FIB-SEM) and atomic force microscopy (AFM). The Si loading etching effect is characterized by the variation of pattern feature depth concerning feature sizes because smaller features tend to etch more slowly than larger ones due to etchants being more difficult to pass through the smaller hole and byproducts being harder to diffuse out too. Thus, the NIL results obtained from the Si master mold contain different pattern geometries concerning pattern quality and residual photoresist layer thickness. The obtained results are pivotal for NIL for fabricating devices with various geometrical nanostructures as the research field moves towards commercial applications.

Sample preparation and data collection for serial block face scanning electron microscopy of mammalian cell monolayers.

Volume electron microscopy encompasses a set of electron microscopy techniques that can be used to examine the ultrastructure of biological tissues and cells in three dimensions. Two block face techniques, focused ion beam scanning electron microscopy (FIB-SEM) and serial block face scanning electron microscopy (SBF-SEM) have often been used to study biological tissue samples. More recently, these techniques have been adapted to in vitro tissue culture samples. Here we describe step-by-step protocols for two sample embedding methods for in vitro tissue culture cells intended to be studied using SBF-SEM. The first focuses on cell pellet embedding and the second on en face embedding. En face embedding can be combined with light microscopy, and this CLEM workflow can be used to identify specific biological events by light microscopy, which can then be imaged using SBF-SEM. We systematically outline the steps necessary to fix, stain, embed and image adherent tissue culture cell monolayers by SBF-SEM. In addition to sample preparation, we discuss optimization of parameters for data collection. We highlight the challenges and key steps of sample preparation, and the consideration of imaging variables.

The impact of macrophage phenotype and heterogeneity on the total internalized gold nanoparticle counts.

Macrophages play a pivotal role in the internalization and processing of administered nanoparticles (NPs). Furthermore, the phagocytic capacity and immunological properties of macrophages can vary depending on their microenvironment, exhibiting a spectrum of polarization states ranging from pro-inflammatory M1 to anti-inflammatory M2. However, previous research investigating this phenotype-dependent interaction with NPs has predominantly relied on semi-quantitative techniques or conventional metrics to assess intracellular NPs. Here, we focus on the interaction of human monocyte-derived macrophage phenotypes (M1-like and M2-like) with gold NPs (AuNPs) by combining population-based metrics and single-cell analysis by focused ion beam-scanning electron microscopy (FIB-SEM). The multimodal analysis revealed phenotype-dependent response and uptake behavior differences, becoming more pronounced after 48 hours. The study also highlighted phenotype-dependent cell-to-cell heterogeneity in AuNPs uptake and variability in particle number at the single-cell level, which was particularly evident in M2-like macrophages, which increases with time, indicating enhanced heteroscedasticity. Future efforts to design NPs targeting macrophages should consider the phenotypic variations and the distribution of NPs concentrations within a population, including the influence of cell-to-cell heterogeneity. This comprehensive understanding will be critical in developing safe and effective NPs to target different macrophage phenotypes.

Evaluation of ER;CR:YSGG Laser and Remineralization Agents on Mineral Density and Ion Levels of Primary and Permanent Enamel.

Objective: The aim of this study is to evaluate the efficacy of two different fluoride varnishes used alone or in combination with laser treatment on permanent and primary tooth enamel. Methods: Ninety-six primary and 96 permanent molar samples were divided into six groups. The levels of calcium, phosphorus, fluoride, and silver ions of each sample were analyzed using energy-dispersive X-ray spectroscopy (EDS). Six different treatments were applied to 12 different groups (n = 15) as control (g1/G1), fluoride varnish (g2/G2), casein phosphopeptide-amorphous calcium phosphate (CPP-ACP)-fluoride varnish (g3/G3), laser (g4/G4), laser+fluoride varnish (g5/G5), and laser+CPP-ACP-fluoride varnish (g6/G6). After the procedures, ion levels were reanalyzed with EDS. The teeth were subjected to the artificial caries-forming procedure and ion levels were again evaluated by EDS. One sample from each group was prepared separately for the focused ion beam-scanning electron microscope measurement; initial and final images were recorded. The obtained data were statistically analyzed with the SPSS 23.0 program. Results: Compared with the initial measurement, phosphorus percentages increased in most of the groups in the last measurement. Calcium percentages of primary teeth increased in the last measurement, except for the g1 group, but in permanent teeth, there was an increase only in the G6 group. There was a statistically significant difference between g1/G1 and g6/G6 groups in the last measurement of phosphorus and calcium percentages. Conclusions: The combined use of laser with CPP-ACP-fluoride varnish enhanced remineralization in the primary and permanent teeth. However, in permanent teeth, the use of laser alone was not as effective as in primary teeth. Therefore, combined usage with CPP-ACP-fluoride varnish can enhance its efficacy. This in vitro study was approved by the local ethics committee of Hacettepe University (Project No.: GO 20/441).

Pore development characteristics and controlling factors of Lower Cambrian Qiongzhusi shale gas reservoir in Sichuan Basin, China

Recently, the shale gas exploration of Lower Cambrian Qiongzhusi Formation in Sichuan Basin has made a great breakthrough. However, the pore characteristics and controlling factors of the shale-gas reservoir in the Qiongzhusi Formation are not clear, which hinders further exploration and development of shale gas. This paper focuses on the Qiongzhusi Formation in the Lower Cambrian of the Sichuan Basin. On the basis of mineral composition and the lithofacies types of shale-gas reservoirs in Qiongzhusi Formation were classified by microscopic observation. Additionally, focused ion beam-scanning electron microscope (FIB-SEM), advanced mineral identification and characterization system (AMICS) analysis, large-field stitching scanning electron microscope technology (MAPS) and nitrogen adsorption experiment were employed to comprehensively determine the differential characteristics and controlling factors of pore development. The results indicate that the pore types in Qiongzhusi Formation are mainly composed of interparticle pores and intercrystalline pores. The pore structure of Qiongzhusi Formation is complex. Significant differences in pore structure parameters are observed among different lithofacies. Quartz and feldspar contribute to the preservation of primary pores due to their strong compressive resistance. Clay minerals and carbonate minerals impede pore development by occluding pores. The influence of organic matter on the pore development is relatively small. According to the mineral composition and pore parameters, laminated shale is a relatively favorable reservoir facies among different lithofacies types.

Electrochemical, Microstructural and Mechanical Investigations of H2O2-Driven Degradation of Ti6Al4V

Titanium and its alloys consitute one of the main classes of materials utilized as non-biodegradable implants in the human body. The corrosion resistance of titanium is mainly due to the presence of a thin, compact and passive layer on its surface. However, this layer degrades in the presence of reactive oxygen species, such as oxygen peroxide (H2O2) [1,2]. This leads to the device degradation and the release of its constituents in the surrounding tissues, which may bring about serious health issues (peri-implantitis, osteolysis, neurotoxicity...) [3]. H2O2 is produced by the immune system during inflammatory episodes by specific enzymes, such as NADPH oxidase and superoxide dismutase [4]. It is also utilized by surgeons at high concentrations (ca. 1 M) during peri-implant tissue disinfection. Ti6Al4V (titanium grade 5, an α + β alloy) is a popular implant material because of its excellent mechanical properties. However, in recent years, Ti6Al4V with equiaxed α grains (and β phase located at the grain boundaries) was reported to undergo a significant degradation by H2O2 characterized by the growth of a thick oxide layer on the α grains and the development of porosity and cracks in the β phase [1]. Ti6Al4V could be susceptible to stress corrosion cracking (SCC) under mechanical load due to the β phase dissolution in H2O2-containing physiological solutions (Figure 1). However, no clear link has been established yet between the corrosive environment, the mechanical load and the microstructure of the Ti6Al4V alloy.This contribution will report about the influence of H2O2 concentration on the in vitro corrosion of Ti6Al4V simulating inflammation conditions (low concentration) as well as disinfection procedures (high concentrations). Results on the electrochemical behavior (monitored by impedance spectroscopy), the degraded microstructure (obtained with focused ion beam scanning electron microscopy and transmission electron microscopy) and the impact of the H2O2-driven degradation on the mechanical properties of the alloy will be presented and discussed.Acknowledgements:The authors gratefully acknowledge the financial support of the French National Research Agency (grant agreement ANR-22-CE93-0007-02) and that of the Swiss National Science Foundation (grant agreement 200021L_213161).

(Invited) Dry Process for Thick Electrode in Batteries

Due to its high operating voltage and cobalt-free chemistry, the spinel-type LiNi0.5Mn1.5O4 (LNMO) cathode material has attracted great interest as one of the few next-generation candidates capable of addressing this combination of challenges. However, severe capacity degradation and poor interphase stability have thus far impeded the practical application of LNMO. In this study, by leveraging a dry electrode coating process, we demonstrate LNMO electrodes with stable full cell operation and ultra-high loading. This excellent cycling stability is ascribed to a stable cathode-electrolyte interphase for charge transfer and ion transport, a highly distributed and interconnected electronic percolation network, and robust mechanical properties. High-quality images collected using plasma focused ion beam scanning electron microscopy (PFIB-SEM) provide additional insight into this behavior, with a complementary 2-D model illustrating how the electronic percolation network in the dry-coated electrodes more efficiently supports homogeneous electrochemical reaction pathways. These results strongly motivate that LNMO as a high voltage cobalt-free cathode chemistry combined with an energy-efficient dry electrode coating process opens the possibility for sustainable electrode manufacturing of cost-effective and high-energy-density cathode materials.

The Mg Electrode Cycling Mechanism in Simple Salt Glyme Electrolytes

The lithium-ion battery (LIB) has been at the forefront of innovations in energy storage since its commercialisation over 30 years ago. As we reach the practical specific capacity limit of current state-of-the-art materials, novel and innovative battery technologies are required to satiate the requirements of energy intensive applications such as electric vehicles. 1 The use of a lithium metal negative electrode has been proposed to increase the energy density, however the high reactivity causes poor cycling performance due to electrolyte degradation, dendrite formation and safety issues. 2 The magnesium electrode offers a solution that has double the theoretical volumetric capacity of the lithium electrode while simultaneously mitigating dendritic growth, reducing raw material costs and greatly increasing sustainability. 3,4 Previous work has shown that traditional LIB electrolytes are not compatible with magnesium metal, and the plating and stripping mechanism that underpins how magnesium electrodes cycle is poorly understood. 5 Here we explored the mechanism of magnesium plating and stripping in glyme-based electrolytes with a range of additional additives. We show that in the pure electrolyte solution, interphase formation is critical to stable cycling but that this is accompanied by significant degradation and accumulation of inactive Mg.6 We establish the three-dimensional chemical composition of Mg deposits in the different electrolyte formulations using a combination of focused ion beam-scanning electron microscopy (FIB-SEM), energy dispersive x-ray spectroscopy (EDX) and transmission electron microscopy (TEM). We use these findings to develop a mechanistic understanding that explains the electrochemical cycling performance of magnesium metal negative electrodes. In situ electrochemical quartz crystal microbalance (EQCM) measurements are used to evaluate the plating and stripping efficiency of the electrodes and to identify non-electrochemical degradation reactions between Mg and the electrolyte solutions.

Quantitative subcellular reconstruction reveals a lipid mediated inter-organelle biogenesis network.

The structures and functions of organelles in cells depend on each other but have not been systematically explored. We established stable knockout cell lines of peroxisomal, Golgi and endoplasmic reticulum genes identified in a whole-genome CRISPR knockout screen for inducers of mitochondrial biogenesis stress, showing that defects in peroxisome, Golgi and endoplasmic reticulum metabolism disrupt mitochondrial structure and function. Our quantitative total-organelle profiling approach for focussed ion beam scanning electron microscopy revealed in unprecedented detail that specific organelle dysfunctions precipitate multi-organelle biogenesis defects, impair mitochondrial morphology and reduce respiration. Multi-omics profiling showed a unified proteome response and global shifts in lipid and glycoprotein homeostasis that are elicited when organelle biogenesis is compromised, and that the resulting mitochondrial dysfunction can be rescued with precursors for ether-glycerophospholipid metabolic pathways. This work defines metabolic and morphological interactions between organelles and how their perturbation can cause disease.

MUSCLE UP: WHAT AGING LOOKS LIKE FROM THE INSIDE OUT

Abstract The Longitudinal Studies Section of the Translational Gerontology Branch of the National Institute on Aging Intramural Research Program has the mission and opportunity to examine aging physiology from the cellular to the full body functional level in model systems and human populations. This symposium presents ongoing research that targets pathologic and normative change in muscle with aging and the significance of these changes for understanding the aging process. Data come from the Baltimore Longitudinal Study of Aging (BLSA) and the Genetic and Epigenetic Signatures of Translational Aging Laboratory Testing (GESTALT) study. In nearly 500 BLSA participants aged 50+, Zenobia Moore evaluates the association between multiple epigenetic age metrics and change over 6 years in mid-femur areas of muscle and subcutaneous and intramuscular fat derived from computed tomography. Qu (Teresa) Tian investigates the relationship between skeletal muscle mitochondrial function and risk of MCI or dementia over 3.3 years in 460 initially cognitively normal BLSA participants and PET and blood biomarkers of AD in available subsamples. In 82 men and women from GESTALT, Stefano Donega examines relationships between physical activity, peak oxygen consumption and muscle oxidative capacity using p31 MR spectroscopy and gene expression, transcript usage and splicing events derived from vastus lateralis biopsies. Using volumetric Focused Ion Beam Scanning Electron Microscopy, Lisa Hartnell shares 3D images of muscle samples at the level of the mitochondrial reticulum and sarcomere from young and old healthy and frail GESTALT participants. Together, these findings highlight the value of muscle as a window on aging.

Mid-Infrared (MIR) Complex Refractive Index Spectra of Polycrystalline Copper-Nitride Films by IR-VASE Ellipsometry and Their FIB-SEM Porosity

Copper-nitride (Cu3N) semiconductor material is attracting much attention as a potential, next-generation thin-film solar light absorber in solar cells. In this communication, polycrystalline covalent Cu3N thin films were prepared using reactive-RF-magnetron-sputtering deposition, at room temperature, onto glass and silicon substrates. The very-broadband optical properties of the Cu3N thin film layers were studied by UV-MIR (0.2–40 μm) ellipsometry and optical transmission, to be able to achieve the goal of a low-cost absorber material to replace the conventional silicon. The reactive-RF-sputtered Cu3N films were also investigated by focused ion beam scanning electron microscopy and both FTIR and Raman spectroscopies. The less dense layer was found to have a value of the static refractive index of 2.304, and the denser film had a value of 2.496. The iso-absorption gap, E04, varied between approximately 1.3 and 1.8 eV and could be considered suitable as a solar light absorber.

Segmentation of cellular ultrastructures on sparsely labeled 3D electron microscopy images using deep learning.

Focused ion beam-scanning electron microscopy (FIB-SEM) images can provide a detailed view of the cellular ultrastructure of tumor cells. A deeper understanding of their organization and interactions can shed light on cancer mechanisms and progression. However, the bottleneck in the analysis is the delineation of the cellular structures to enable quantitative measurements and analysis. We mitigated this limitation using deep learning to segment cells and subcellular ultrastructure in 3D FIB-SEM images of tumor biopsies obtained from patients with metastatic breast and pancreatic cancers. The ultrastructures, such as nuclei, nucleoli, mitochondria, endosomes, and lysosomes, are relatively better defined than their surroundings and can be segmented with high accuracy using a neural network trained with sparse manual labels. Cell segmentation, on the other hand, is much more challenging due to the lack of clear boundaries separating cells in the tissue. We adopted a multi-pronged approach combining detection, boundary propagation, and tracking for cell segmentation. Specifically, a neural network was employed to detect the intracellular space; optical flow was used to propagate cell boundaries across the z-stack from the nearest ground truth image in order to facilitate the separation of individual cells; finally, the filopodium-like protrusions were tracked to the main cells by calculating the intersection over union measure for all regions detected in consecutive images along z-stack and connecting regions with maximum overlap. The proposed cell segmentation methodology resulted in an average Dice score of 0.93. For nuclei, nucleoli, and mitochondria, the segmentation achieved Dice scores of 0.99, 0.98, and 0.86, respectively. The segmentation of FIB-SEM images will enable interpretative rendering and provide quantitative image features to be associated with relevant clinical variables.

Detection of atmospheric aerosols and terrestrial nanoparticles collected in a populous city in southern Brazil.

The main objective of this study is to analyze hazardous elements in nanoparticles (NPs) (smaller than 100nm) and ultrafine particles (smaller than 1µm) in Porto Alegre City, southern Brazil using a self-made passive sampler and Sentinel-3B SYN satellite images in 32 collection points. The Aerosol Optical Thickness proportion (T550) identification was conducted using images of the Sentinel-3B SYN satellite at 634 points sampled in 2019, 2020, 2021, and 2022. Focused ion beam scanning electron microscopy analyses were performed to identify chemical elements present in NPs and ultrafine particles, followed by single-stage cascade impactor to be processed by high-resolution transmission electron microscopy. This process was coupled with energy-dispersive X-ray spectroscopy and later analysis via secondary ion mass spectrometry. Data was acquired from Sentinel-3B SYN images, normalized to a standard mean of 0.83µg/mg, at moderate spatial resolution (260m), and modeled in the Sentinel Application Platform (SNAP) software v.8.0. Statistical matrix data was generated in the JASP software (Jeffreys's Amazing Statistics Program) v.0.14.1.0 followed by a K-means cluster analysis. The results demonstrate the presence of between 1 and 100nm particles of the following chemical elements: Si, Al, K, Mg, P, and Ti. Many people go through these areas daily and may inhale or absorb these elements that can harm human health. In the Sentinel-3B SYN satellite images, the sum of squares in cluster 6 is 168,265 and in cluster 7 a total of 21,583. The use of images from the Sentinel-3B SYN satellite to obtain T550 levels is of great importance as it reveals that atmospheric pollution can move through air currents contaminating large areas on a global scale.

Bundling of collagen fibrils influences osteocyte network formation during bone modeling

Osteocytes form a cellular network by gap junctions between their cell processes. This network is important since intercellular communication via the network is essential for bone metabolism. However, the factors that influence the formation of this osteocyte network remain unknown. As the early stage of osteocyte network formation occurs on the bone surface, we observed a newly formed trabecular bone surface by orthogonal focused ion beam-scanning electron microscopy. The embedding late osteoblast processes tended to avoid bundled collagen fibrils and elongate into sparse collagen fibrils. Then, we examined whether the inhibition of bundling of collagen fibrils using a potent lysyl oxidase inhibitor, β-aminopropionitrile (BAPN) changed the cellular network of the chick calvaria. The osteocyte shape of the control group was spindle-shape, while that of the BAPN group was sphere-shaped. In addition, the osteocyte processes of the control group were elongated vertically to the long axis of the cell body, whereas the osteocyte processes of the BAPN group were elongated radially. Therefore, it was suggested that the bundling of collagen fibrils influences normal osteocyte network formation during bone modeling.

Size and shape analysis of micro- to nano-particles of quartz powders using advanced electron microscopy and laser diffraction methods

The paper deals with particle analysis and the evaluation of the shape and size distribution of micro- to nano-sized quartz particles as a key parameter for the evaluation of quality in many industrial fields, particularly for high-purity applications. Samples are analyzed by laser diffraction, the most commonly used method for determining the size of the particles; for precise measurement with the possibility of high-resolution imaging of particles, scanning electron microscopy was applied. For morphological and size analysis of electrically non-conductive micro- to nano-particles under environmentally compatible conditions of elevated gas pressure and humidity, environmental scanning electron microscopy was selected as the most suitable. Moreover, the high-end methods of focused ion beam scanning electron microscopy and serial block-face scanning electron microscopy enabled the analysis of particles in the epoxy resin volume including spatial imaging. The application of image post-processing as an important part of the electron microscopy workflow is also discussed.

Characterization for 3D pore-fracture systems in Hot-Dry-Rock samples from Gonghe Basin, China

As a new type of renewable energy, Hot-Dry-Rock (HDR) is gaining increasing attention. The physical properties of rock influenced by the pore-fracture structure are vital for the hydraulic and chemical characteristics of HDR reservoirs. However, the natural pore-fracture structure in HDR has not been sufficiently studied. To fulfill this gap, we used focused ion beam-scanning electron microscopy (FIB-SEM) to construct three-dimensional (3D) digital models for four granite samples from the deep HDR reservoir in Gonghe Basin, Qinghai Province, China, and obtained the nanoscale structure of the pores and fractures in HDR reservoir for the first time. Our analysis based on the 3D models indicates that the heat transfer area in the HDR reservoir is mainly from fractures and crack-like pores, but a large amount of small but regular pores could be neglected when heat transfer in HDR is considered. The permeability of HDR reservoirs at nanoscale varies from 9.414 nD to 0.082 mD, and the high permeability in this study is accompanied by throughgoing fractures. The variation of porosity and permeability of deep HDR at nanoscale in this study is much more severe than that of shallow granites previously studied at microscale. The performance of geothermal field is significantly influenced by the size of throats and the average size and tortuosity of the interconnected pore-fracture network in HDR. The hydraulic aperture could be 32-58% lower than the mechanical aperture of the natural fractures in HDR reservoirs, which is a vital clue for the numerical simulation associated with enhanced geothermal systems.