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Keiichi Moriguchi, Takamichi Jogahara, Senichi Oda, Masaki Honda
A 3, 3'-diaminobenzidine (DAB)-based method was used to detect the localization of endogenous peroxidase activity in Indian rhinoceros (Rhinoceros unicornis) parotid gland acinar cells. The tissue had previously been resin-embedded in gelatin capsules for routine electron microscopic observations and thus pre-incubation for endogenous peroxidase analysis was not possible. We attempted to demonstrate the relationship between secretory granules (SGs) in resin ultrathin sections of Indian rhinoceros parotid gland acinar cells and endogenous peroxidase activity...
October 31, 2018: Microscopy
Yuka Sato, Shigeo Okabe
Circuit-level alternations in patients of autism spectrum disorder (ASD) is under active investigation and detailed characterization of synapse morphology in ASD model mice should be informative. We utilized focused ion beam milling and scanning electron microscopy (FIB-SEM) to obtain three-dimensional images of synapses in the layer 2/3 of the somatosensory cortex from a mouse model for ASD with human 15q11-13 chromosomal duplication (15q dup mice). We found a trend of higher spine density and a higher fraction of astrocytic contact with both spine and shaft synapses in 15q dup mice...
October 29, 2018: Microscopy
Tomohiro Kubo, Toshiyuki Oda
The diversity of α- and β-tubulin is facilitated by various post-translational modifications (PTMs), such as acetylation, tyrosination, glycylation, glutamylation, phosphorylation and methylation. These PTMs affect the stability and structure of microtubules as well as the interaction between microtubules and microtubule-associated proteins, including molecular motors. Therefore, it is extremely important to investigate the roles of tubulin PTMs for understanding the cell cycle, cell motility and intracellular trafficking...
October 26, 2018: Microscopy
Shunichi Morikawa, Azusa Sato, Taichi Ezaki
Although conventional toluidine blue staining is a common technique used for rapid observation of semithin sections prior to transmission electron microscopy, it is monochromatic and insufficient for accurate identification of different tissue components by light microscopy. Additionally, polychromatic staining methods generally require step-by-step processes involving different dyes, and it is often difficult to balance the color tone of each step. In this study, we developed a simple polychromatic staining method for epoxy-embedded tissue sections...
October 15, 2018: Microscopy
Yoshifumi Oshima
Recently, visualization of lithium ions has brought great benefit for understanding lithium movement in the cathode, electrolyte and anode materials of lithium ion battery. It has been achieved by several methods such as spherical aberration corrected scanning transmission electron microscopy, multivariable analysis, advanced electron holography and so on, which was reviewed by the special issue, 'Challenges for Lithium Detection' in Microscopy (Vol. 66, No. 1, 2017). In this paper, recent research progress in lithium detection is introduced as a follow-up for the special issue...
October 15, 2018: Microscopy
Noriyuki Inoue, Yoshiko Takashima, Mitsuo Suga, Toshiaki Suzuki, Yoshikazu Nemoto, Osamu Takai
Wet specimens are notoriously difficult to image in scanning electron microscopes (SEM) owing to evaporation from the required vacuum of the specimen chamber. Traditionally, this issue has been addressed by increasing the specimen chamber pressure. Unfortunately, observation under high specimen chamber pressure cannot prevent the initial evaporation effects. The wet cover method, where the original surface water is retained (and, therefore, considered wet), provides a way to introduce and subsequently image specimens that are sensitive to evaporation within a SEM, while preventing evaporation-related damage, and to observe interesting specimen-water interactions...
October 11, 2018: Microscopy
Wataru Yashiro
The self-imaging phenomenon referred to as the Talbot effect in the field of optics was discovered by H.F. Talbot in the 1830s, and is now widely used for imaging using not only visible light but also X-rays, electrons, neutrons, and matter waves. In this review, the author introduces the current progress being made in hard-X-ray imaging microscopy based on the self-imaging phenomenon. Hard-X-ray imaging microscopy is a promising technique for non-destructively visualizing internal structures in specimens with a spatial resolution up to a few tens of nanometers...
October 11, 2018: Microscopy
Wanxiang Qin, Min Luo, Kejian Wang, Mei Yang, Huajun Sheng, Guiqiong He
The autofluorescence of animal tissues complicates the results obtained using fluorescence microscopy. Many techniques have been used to reduce autofluorescence; however, all these techniques have the disadvantage of reducing the intensity of immunofluorescence staining. We observed the features of autofluorescence in formalin-fixed paraffin-embedded (FFPE) vascularized liver and kidney sections and assessed the effects of an intravascular treatment with erythrocyte lysis solution (ELS) before a routine perfusion with normal saline (NS) and Sudan Black B (SBB) treatment after antigen retrieval on reducing autofluorescence reduction and the visualization of antigens to establish an optimal method for reducing autofluorescence...
October 11, 2018: Microscopy
Sidney L Shaw, David Thoms, James Powers
The advent of super-resolution techniques in biological microscopy has opened new frontiers for exploring the molecular distribution of proteins and small molecules in cells. Improvements in optical design and innovations in the approaches for the collection of fluorescence emission have produced substantial gains in signal from chemical labels and fluorescent proteins. Structuring the illumination to elicit fluorescence from specific or even random patterns allows the extraction of higher order spatial frequencies from specimens labeled with conventional probes...
October 8, 2018: Microscopy
Yuji Konyuba, Tomohiro Haruta, Yuta Ikeda, Tomohisa Fukuda
In transmission electron microscopy (TEM), silicon nitride (SiN) films are widely used as sample-supporting films owing to their robustness. We fabricated large-scale SiN films deposited by low-pressure chemical vapor deposition (LPCVD). This preparation method is advantageous for large window areas, since it yields films with control over properties such as tension and thickness. We fabricated large SiN windows for mounting large ultrathin sections and for acquiring large-area TEM images. Thus, sample sections sliced by conventional sample preparation techniques were successfully mounted on these sample-supporting films...
September 29, 2018: Microscopy
Ryosuke Okamoto, Kazuhide Hayashi, Satoshi Matsumoto, Naomi Suzuki, Masami Terauchi, Kenji Tsuda
LiMn1.5Ni0.5O4 is an excellent candidate as a cathode-active material in high-voltage lithium-ion batteries and studied using atomic resolution scanning transmission electron microscope. High-angle annular dark-field (HAADF) images obtained at [100] orientation demonstrate that Mn and Ni atoms are regularly ordered at octahedral sites in a spinel structure, in a 3:1 ratio between columns with high and low intensities. Simulations of HAADF images revealed that atomic columns including Mn exhibit a larger intensity than that by Ni columns, primarily because of the effect of the Debye-Waller factor...
October 1, 2018: Microscopy
Pilar Cossio, Matteo Allegretti, Florian Mayer, Volker Müller, Janet Vonck, Gerhard Hummer
The 'Bayesian inference of electron microscopy' (BioEM) framework makes it possible to determine the stoichiometry of protein complexes using 3D coarse-grained models and a relatively small number of cryo-electron microscopy images as input. We applied the method to determine the most probable rotor ring stoichiometry of the archaeal Na+ ATP synthase from Pyrococcus furiosus, a multisubunit complex able to produce ATP under extreme conditions. Archaeal ATP synthases consist of a catalytic A1 part and a membrane-embedded AO portion...
October 1, 2018: Microscopy
Jinfeng Yang, Yoichi Yoshida, Hidehiro Yasuda
Ultrafast electron microscopy (UEM) with femtosecond temporal resolution is a 'dream machine' that has been long envisioned for the study of ultrafast structural dynamics in materials. For this purpose, we developed a prototype UEM with relativistic femtosecond electron pulses generated by a radio-frequency acceleration-based photoemission gun. TEM images of polystyrene latex particles and gold nanoparticles were observed using approximately 100-fs-long electron pulses with energies of 3.1 MeV. The effect of emittance and the number of pulses on the images were investigated...
October 1, 2018: Microscopy
Tetsuya Akashi, Yoshio Takahashi, Ken Harada, Toshio Onai, Yoshimasa A Ono, Hiroyuki Shinada, Yasukazu Murakami
The coherency of a 1.2-MV transmission electron microscope was evaluated through illumination semiangles calculated from lengths over which Fresnel fringes can be observed. These lengths were determined from the diameters of circular holes fully filled with Fresnel fringes, i.e. this method allows lengths to be accurately measured even if micrographs are subjected to distortions. The smallest illumination semiangle of 4.0 × 10-9 rad was obtained for a circular hole with a diameter of 191 μm. In addition, electron beam brightness was estimated to be approximately 3 × 1014 A/m2·sr from the obtained illumination semiangle values and current densities...
October 1, 2018: Microscopy
Kohei Tandokoro, Masayasu Nagoshi, Takashi Kawano, Kaoru Sato, Katsushige Tsuno
Scanning electron microscopy (SEM) is a powerful tool for observing the surface of materials. Modern SEM systems have multiple detectors with different geometries. Consequently, the SEM image contrast depends on the instrument and experimental conditions even for the same sample. Understanding the SEM imaging mechanism is necessary to clarify SEM contrast. In this paper, low-voltage (LV)-SEM image contrast is investigated by comparing LV-SEM images and electron trajectory simulation results. Surface observations of oxides on a steel surface, positive charging contrast and topographic contrast in the image systematically changed with the working distance (WD)...
October 1, 2018: Microscopy
Taiji Yamashita, Kenji Miyamoto, Hitoshi Yonenobu
A new pretreatment method using room-temperature ionic liquid (IL) was proposed for observing wood specimens in scanning electron microscopy (SEM). A variety of concentrations were examined for ethanol solution of the IL, [Emim][MePO3Me], to determine an optimal pretreatment procedure. It was concluded that 10% ethanol solution of the IL was the most adequate to acquire good SEM images. Using the procedure optimized, SEM images were taken for typical anatomical types of modern soft and hardwood species and archeological wood...
October 1, 2018: Microscopy
Takeshi Higuchi, Takashi Gondo, Hiroya Miyazaki, Akemi Kumagai, Keizo Akutagawa, Hiroshi Jinnai
An in situ straining holder capable of tensile deformation and high-angle tilt for electron tomography was developed for polymeric materials. The holder has a dedicated sample cartridge, on which a variety of polymeric materials, such as microtomed thin sections of bulk specimens and solvent-cast thin films, can be mounted. Fine, stable control of the deformation process with nanoscale magnification was achieved. The holder allows large tensile deformation (≃800 μm) with a large field of view (800 × 200 μm before the deformation), and a high tilt angle (±75°) during in situ observations...
October 1, 2018: Microscopy
Junko Matsuda, Tatsuya Kawasaki, Shotaro Futamura, Tsutomu Kawabata, Shunsuke Taniguchi, Kazunari Sasaki
In situ transmission electron microscopy (TEM) observations of a Ni(O)-Sc2O3-stabilized ZrO2 (ScSZ; 10 mol% Sc2O3, 1 mol% CeO2, 89 mol% ZrO2) anode in a solid oxide fuel cell (SOFC) have been performed at high temperatures under a hydrogen/oxygen gas atmosphere using an environmental transmission electron microscope (ETEM); the specimens were removed from cross-sections of the real SOFC by focused ion beam milling and lifting. When heating the NiO-ScSZ anode under a hydrogen atmosphere of 3 mbar in ETEM, nano-pores were formed at the grain boundaries and on the surface of NiO particles at around 400°C due to the volume shrinkage accompanying the reduction of NiO to Ni...
October 1, 2018: Microscopy
Yuta Yamamoto, Mizuki Hattori, Junya Ohyama, Atsushi Satsuma, Nobuo Tanaka, Shunsuke Muto
In this article, we demonstrate that a convolutional neural network (CNN) can be effectively used to determine the presence of twins in the atomic resolution scanning transmission electron microscopy (STEM) images of catalytic Au nanoparticles. In particular, the CNN screening of Hough transformed images resulted in significantly higher accuracy rates as compared to those obtained by applying this technique to the raw STEM images. The proposed method can be utilized for evaluating the statistical twining fraction of Au nanoparticles that strongly affects their catalytic activity...
August 15, 2018: Microscopy
Daisuke Koga, Satoshi Kusumi, Tsuyoshi Watanabe
Scanning electron microscopes have longer focal depths than transmission electron microscopes and enable visualization of the three-dimensional (3D) surface structures of specimens. While scanning electron microscopy (SEM) in biological research was generally used for the analysis of bulk specimens until around the year 2000, more recent instrumental advances have broadened the application of SEM; for example, backscattered electron (BSE) signals under low accelerating voltages allow block-face and section-face images of tissues embedded in resin to be acquired...
June 15, 2018: Microscopy
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