Microscopy

Electron channeling contrast imaging (ECCI) was applied by precisely controlling the 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...

Hydrogen is attracting attention as an energy carrier for realizing a low-carbon society, because it can directly convert the energy obtained from chemical reactions into electrical energy without carbon dioxide emissions. This paper presents in situ transmission electron microscopic (TEM) observations related to hydrogen storage in metal and metal hydrides, hydrogen embrittlement of metallic materials used for storing and transporting hydrogen in containers and pipes, and fuel cells and water electrolysis using metal catalysts and oxides as electrode materials...

Morphogenesis is a developmental process that shapes multicellular organisms through complex and cooperative cellular movements. To understand the complex interplay between genetic programs and resulting multicellular morphogenesis, it is essential to characterize the morphologies and dynamics at the single-cell level, with an understanding of how physical forces serve as both signaling components and driving forces of tissue deformations. In recent years, advances in microscopy techniques have led to improvements in imaging speed, resolution, and depth...

Solid-state batteries have potential to realize a rechargeable Li metal anode. However, several challenges persist in the charging and discharging processes of the Li metal anode, which require a fundamental understanding of Li plating and stripping across the interface of solid-state electrolytes to address. This review overviews studies on Li metal anodes in solid-state batteries using in situ observation techniques with an emphasis on Li electrodeposition and dissolution using scanning electron microscopy (SEM) and solid electrolytes (SEs) such as lithium phosphorus oxynitride (LiPON) and garnet-type compounds such as Li7La3Zr2O12 (LLZ)...

In situ transmission/scanning transmission electron microscopy (TEM/STEM) measurements have taken a central stage for establishing structure-chemistry-property relationship over the past couple of decades. The challenges for realizing 'a lab-in-gap', i.e., gap between the objective lens pole-pieces, or 'a lab-on-chip', to be used to carry out experiments are being met through continuous instrumental developments. Commercially available TEM columns and sample holder that have been modified for in situ experimentation have contributed to uncovering structural and chemical changes occurring in the sample when subjected external stimulus such as temperature, pressure, radiation (photon, ions, electrons), environment (gas, liquid, magnetic or electrical field), or a combination thereof...

During the in-situ TEM observations, the diverse functionalities of different specimen holders play a crucial role. We hereby provide a comprehensive overview of the main types of holders, associated technologies, and case studies pertaining to the widely employed heating and gas heating methods, from their initial developments to the latest advancement. In addition to the conventional approaches, we also discuss the emergence of holders that incorporate a micro-electro-mechanical system (MEMS) chip for in-situ observations...

Quantifying the number of molecules from fluorescence microscopy measurements is an important topic in cell biology and medical research. In this work, we present a consecutive algorithm for super-resolution (stimulated emission depletion (STED)) scanning microscopy that provides molecule counts in automatically generated image segments and offers statistical guarantees in form of asymptotic confidence intervals. To this end, we first apply a multiscale scanning procedure on STED microscopy measurements of the sample to obtain a system of significant regions, each of which contains at least one molecule with prescribed uniform probability...

We report direct observation by electron holography of the spin polarization of electrons in a vacuum region around a charged SiO2 wire coated with Pt-Pd. Irradiating the SiO2 wire with 300-keV electrons caused the wire to become positively charged due to the emission of secondary electrons. The spin polarization of these electrons interacting with the charged wire was observed in situ using a phase reconstruction process under an external magnetic field. The magnetic field of the spin-polarized electrons was simulated taking into account the distribution of secondary electrons and the effect of the external magnetic field...

The two-dimensional observation of ultrathin sections from resin-embedded specimens provides insufficient understanding of the three-dimensional (3D) morphological information of membranous organelles. The osmium maceration method, developed by Professor Tanaka's group over 40 years ago, is the only technique that allows direct observation of the 3D ultrastructure of membrane systems using scanning electron microscopy (SEM), without the need for any reconstruction process. With this method, the soluble cytoplasmic proteins are removed from the freeze-cracked surface of cells while preserving the integrity of membranous organelles, achieved by immersing tissues in a diluted osmium solution for several days...

High-speed atomic force microscopy is now a widely used technique to study dynamics of single biomolecules and complex structures. In the past, it has mainly been used to capture surface topography as structural analysis, leading to important discoveries not attainable by other methods. Similar to conventional atomic force microscopy, the scope of high-speed atomic force microscopy was recently expanded to encompass quantities beyond topography, such as the measurement of mechanical properties. This review delves into various methodologies for assessing mechanical properties, ranging from semi-quantitative approaches to precise force measurements and their corresponding sample responses...

We present a graph neural network (GNN)-based framework applied to large-scale microscopy image segmentation tasks. While deep learning models, like convolutional neural networks (CNNs), have become common for automating image segmentation tasks, they are limited by the image size that can fit in the memory of computational hardware. In a GNN framework, large-scale images are converted into graphs using superpixels (regions of pixels with similar color/intensity values), allowing us to input information from the entire image into the model...

The advent of single-molecule atomic-resolution time-resolved electron microscopy (SMART-EM) has created a new field of "cinematic chemistry," allowing for the cinematographic recording of dynamic behaviors of organic and inorganic molecules and their assembly. However, the limited electron dose per frame of video images presents a major challenge in SMART-EM. Recent advances in direct electron counting cameras and techniques to enhance image quality through the implementation of a denoising algorithm have enabled the tracking of stochastic molecular motions and chemical reactions with sub-millisecond temporal resolution and sub-angstrom localization precision...

In this study, a new method for the phase retrieval of electron rocking curves observed using convergent-beam electron diffraction, which is applicable to the determination of three-dimensional lattice displacement fields along the beam direction, is proposed. Total variation and total squared variation regularizations are introduced for phase retrieval to suppress overfitting to noise or background signals in the rocking curves and to reproduce the sparse characteristics of displacement fields, which exist only near lattice defects...

Myelin-associated glycoprotein (MAG), released from pre-degenerated distal nerves following axotomy, blocks the regrowth of sprouts and naked axons. Ensheathed axons, however, continue to elongate and reach MAG-releasing distal nerves. To determine the regenerative mechanism of ensheathed axons without navigators of axonal growth cones by the film model method, we inserted a MAG-releasing distal nerve segment treated with liquid nitrogen (N2DS) between two films, facing the proximal end of the common peroneal nerves in mice transected four days earlier for axons to become ensheathed...

We developed an in-situ shear test system suitable for transmission electron microscopy (TEM) observations, which enabled us to examine the shear deformation behaviours inside soft materials at nanoscale resolutions. This study was conducted on a nanoparticle-filled rubber to investigate its nanoscale deformation behaviour under a large shear strain. First, the shear deformation process of a large area in the specimen was accurately examined and proven to exhibit an almost perfect simple shear. At the nanoscale, voids grew along the maximum principal strain during shear deformation...

Nanosized precipitates have a critical role in increasing the strength of metallic alloys. There are many reports that the initial precipitates are metastable phases holding a different composition and crystal structure from the equilibrium precipitate. The metastable precipitate transforms to its stable phase during heat treatment. A transmission electron microscope enables researchers to study the phase transition of metastable precipitates to stable phases due to its fine resolution in identifying crystal structure and chemical composition...

Herein, we review notable points from observations of electrochemical reactions in a liquid electrolyte by liquid-phase electron microscopy (LP-EM). In situ microscopic observations of electrochemical reactions are urgently required, particularly to solve various battery issues. Battery performance is evaluated by various electrochemical measurements of bulk samples. However, it is necessary to understand the physical/chemical phenomena occurring in batteries to elucidate the reaction mechanisms. Thus, in situ microscopic observation is effective for understanding the reactions that occur in batteries...

In recent years, the technique of scanning electron microscopy (SEM) observation with low landing energy of a few keV or less has become common. We have especially focused on the drastic contrast change at near 0 eV. Using a patterned sample consisting of Si, Ni and Pt, threshold energies where total reflection of incident electrons occur was investigated by SEM at near 0 eV. In both the cases of in-situ and ex-situ sample cleaning, drastic changes in the brightness of each material were observed at near 0 eV, with threshold energies in the order Si < Ni < Pt...

We have demonstrated a new data analysis method that enables nondestructive depth profiling of a multilayer thin-film sample from energy-dispersive X-ray spectroscopy (EDX) data without the assumption of initial profiles. This method is based on a quadratic programming problem and allows for three-dimensional elemental mapping in the sample without destroying it, by performing depth profiling for all the pixels in the EDX two-dimensional mapping data. In this paper, first nondestructive depth profiling of two samples with different multilayer structures was performed using the proposed method...

We conducted a comprehensive analysis of the surface microtexture of kefir biofilms grown on Theobroma grandiflorum Shum (cupuaçu) juice using atomic force microscopy. Our goal was to investigate the unique monofractal and multifractal spatial patterns of these biofilms to complement the existing limited literature. The biofilms were prepared dispersing four different concentrations of kefir grains in cupuaçu juice. Our morphological analysis showed that the surface of the obtained biofilms is essentially formed by the presence of cupuaçu fibers and microorganisms like lactobacilli and yeast...