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...
The haploid gametophyte generation of ferns is an excellent experimental material for cell biology studies because of its simple structure and high sensitivity to light. Each step of the developmental process, such as cell growth, cell cycle and the direction of cell division, is controlled, step by step, by light, unlike what happens in complex seed plant tissues. To perform...
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...
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...
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...
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...
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...
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 ‘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...
The recent ‘resolution revolution’ in structural analyses of cryo-electron microscopy (cryo-EM) has drastically changed the research strategy for structural biology. In addition to X-ray crystallography and nuclear magnetic resonance spectroscopy, cryo-EM has achieved the structural analysis of biological molecules at near-atomic resolution, resulting in the Nobel Prize in...
In this review, we introduce digital holographic techniques and recent progress in multidimensional sensing by using digital holography. Digital holography is an interferometric imaging technique that does not require an imaging lens and can be used to perform simultaneous imaging of multidimensional information, such as three-dimensional structure, dynamics, quantitative phase...
What does the diffraction pattern from a single atom look like? How does it differ from the scattering from long-range potential? With the development of new high-dynamic range pixel array detectors to measure the complete momentum distribution, these questions have immediate relevance for designing and understanding momentum-resolved imaging modes. We explore the asymptotic...
Multivariate analysis is a powerful tool to process spectrum imaging datasets of electron energy loss spectroscopy. Most spatial variance of the datasets can be explained by a limited numbers of components. We explore such dimension reduction to facilitate quantitative analyses of spectrum imaging data, supervising the spectral components instead of spectra at individual pixels...
This publication is a systematic investigation of the effect the improvement of dark-reference images has on the resulting bright-field images. For this, data were acquired with three different charge-coupled device cameras attached to two different transmission electron microscopes. Multi-frame acquisitions and methods to correct x-ray noise are introduced and quantified as...
Specimen and stage drift as well as scan distortions can lead to a mismatch between true and desired electron probe positions in scanning transmission electron microscopy (STEM) which can result in both linear and nonlinear distortions in the subsequent experimental images. This problem is intensified in STEM spectrum and diffraction imaging techniques owing to the extended dwell...
Since the development of parallel electron energy loss spectroscopy (EELS), charge-coupled devices (CCDs) have been the default detectors for EELS. With the recent development of electron-counting direct-detection cameras, micrographs can be acquired under very low electron doses at significantly improved signal-to-noise ratio. In spectroscopy, in particular in combination with a...
Electron energy-loss spectroscopy (EELS) has been investigated in the range from 2 to >10 keV using an optimized optical coupling of the microscope to the spectrometer to improve the high loss performance in EELS. It is found that excellent quality data can now be acquired up until about 5 keV, suitable for both energy loss near edge structure (ELNES) studies of oxidation and...
We present the surface plasmon resonance modes in three-dimensional (3D) upright split ring resonators (SRR) as studied by correlative cathodoluminescence (CL) spectroscopy in a scanning electron microscope (SEM) and electron energy loss spectroscopy (EELS) in a transmission electron microscope. We discuss the challenges inherent in studying the plasmon modes of a 3D...
Heterogeneous assemblies of molecules (Rhodamine B) adsorbed onto a nano-corrugated metallic surface (a percolated Au network) are investigated using electron energy loss spectroscopy in the scanning transmission electron microscope (STEM-EELS). Our first measurements target the native metallic substrate, which consists of a commercial Au thin film atop an ultrathin carbon...
Using spatially resolved Electron Energy-Loss Spectroscopy, we investigate the excitation of long-wavelength surface optical vibrational modes in elementary types of nanostructures: an amorphous SiO2 slab, an MgO cube, and in the composite cube/slab system. We find rich sets of optical vibrational modes strongly constrained by the nanoscale size and geometry. For slabs, we find...