Microscopy (Oxford, England)最新文献

The detective quantum efficiency (DQE) is generally accepted as the main figure of merit for the comparison between electron detectors, and most of the time given as a unique number at the Nyquist frequency while it is known to vary with electron dose. It is usually estimated, thanks to a method improved by McMullan in 2009. The purpose of this work is to analyze and to criticize this DQE extraction method on the basis of measurement and model results, and to give recommendations for fair comparison between detectors, wondering if the DQE is the right figure of merit for electron detectors.

We report a novel class of scanning transmission electron microscopy with Hilbert-differential phase contrast (HDP-STEM) that displays nanostructures of thin samples in a topographical manner. A semicircular π-phase plate (PP) was used as an optical device for manipulating electron waves in HDP-STEM. This is the different design from the Zernike PP used in our previous phase plate STEM (P-STEM), but both must be placed in the front focal plane of the condenser lens. HDP-STEM images of multiwalled carbon nanotubes showed higher contrast than those obtained by conventional bright-field STEM. As the PP of the HDP-STEM is nonsymmetrical, several different images were obtained by changing the detection conditions. A two-dimensional electron detector was also used to remove the scattering contrast component in the same way as with the Zernike PP and obtain an image containing only (differential) phase contrast.

Correlative array tomography, combining light and electron microscopy via serial sections, plays a crucial role in the three-dimensional ultrastructural visualization and molecular distribution analysis in biological structures. To address the challenges of aligning fluorescence and electron microscopy images and aligning serial sections of irregularly shaped biological specimens, we developed a diamond notch knife, a new tool for puncturing holes using a diamond needle. The diamond needle featured a triangular and right-angled tip, enabling the drilling of deep holes upon insertion into the polished block face. This study describes the application of the diamond notch knife in correlative array tomography.

The three-dimensional (3D) anatomical structure of living organisms is intrinsically linked to their functions, yet modern life sciences have not fully explored this aspect. Recently, the combination of efficient tissue clearing techniques and light-sheet fluorescence microscopy (LSFM) for rapid 3D imaging has improved access to 3D spatial information in biological systems. This technology has found applications in various fields, including neuroscience, cancer research, and clinical histopathology, leading to significant insights. It allows imaging of entire organs or even whole bodies of animals and humans at multiple scales. Moreover, it enables a form of spatial omics by capturing and analyzing cellome information, which represents the complete spatial organization of cells. While current 3D imaging of cleared tissues has limitations in obtaining sufficient molecular information, emerging technologies such as multi-round tissue staining and super-multicolor imaging are expected to address these constraints. 3D imaging using tissue clearing and light-sheet microscopy thus offers a valuable research tool in the current and future life sciences for acquiring and analyzing large-scale biological spatial information.

X-ray microscopy using computed tomography (CT) is an excellent three-dimensional imaging instrument. Three-dimensional X-ray microscopy (3DXRM) is a nondestructive imaging technique used to inspect internal and external structures in units of submicrometers or less. The 3DXRM, although attractive, is mostly used as an observation instrument and is limited as a measurement system in quantitative evaluation and quality control. Calibration is required for use in measurement systems such as coordinate measurement systems, and specific standard samples and evaluation procedures are needed. The certified values of the standard samples must ideally be traceable to the International System of Units (SI). In the 3DXRM measurement system, line structures (LSs) are fabricated as prototype standard samples to conduct magnification calibration. In this study, we evaluated the LS intervals using calibrated cross-sectional scanning electron microscopy (SEM). A comparison of the evaluation results between SEM and 3DXRM for the LS intervals provided the magnification calibration factor for 3DXRM and validated the LSs, whereby the interval methods and feasibility of constructing an SI traceability system were evaluated using the calibrated SEM. Consequently, a magnification calibration factor of 1.01 was obtained for 3DXRM based on the intervals of the LSs evaluated by SEM. A possible route for realizing SI-traceable magnification calibration of 3DXRM has been presented.

The surface sensitivity of high-resolution secondary electron (SE) imaging is examined using twisted bilayers of MoS2 stacked at an angle of 30-degree. High-resolution SE images of the twisted bilayer MoS2 show a honeycomb structure composed of Mo and S atoms, elucidating the monolayer structure of MoS2. Simultaneously captured annular dark-field scanning transmission electron microscope images from the same region show the projected structure of the two layers. That is, the SE images from the bilayer MoS2 selectively visualize the surface monolayer. It is noted that SE yields from the surface monolayer are approximately 3 times higher than those from the second monolayer, likely attributable to attenuation when SEs emitted from the second layer traverse the surface layer. Mini abstract: The surface sensitivity of atomic resolution secondary electron imaging is examined using MoS2 bilayers, the thinnest system composed of a surface layer and substrate. This study reveals that the secondary electrons visualize the atomic arrangement of the surface monolayer three times more intensely than that of the second layer.

In this study, we experimentally analyzed the charging phenomenon when an insulating resist film on a conductive layer formed on bulk glass is irradiated by electron beams. To quantify the charging potential induced, an electrostatic force microscope device was installed in the scanning electron microscope sample chamber, and potential distributions formed under various exposure conditions were obtained. Based on the results obtained, a model for charge accumulation within the sample, explaining positive and negative charging and their transitions, was developed. At an electron beam acceleration voltage of 30 kV, the following observations were made: "global charging" could be avoided by applying -5V to the sample. Regarding "local charging" near the exposure area of the electron beam, at low exposure doses, emission of secondary electrons from the sample surface induced positive charging, while the accumulation of incident electrons within the sample induced negative charging. At exposure doses where the effects of both are balanced, the sample exhibited zero potential, revealing the appearance of the "first zero-cross exposure dose". At higher exposure doses, the sample transitions from negative to positive as the exposure dose increases due to the electron beam induced conduction, resulting in the so-called "second zero-cross exposure dose". The exposure dose dependence of the charging potential distribution at various acceleration voltages was obtained. In particular, we found that at an acceleration voltage of 0.6 kV, the sample surface is not charged even when exposed to small to very large doses of electron beams.

We have demonstrated localized surface plasmon (LSP)-enhanced cathodoluminescence (CL) from an atomic layer deposition (ALD)-grown Al2O3/ZnO/Al2O3 heterostructure to develop a bright nanometer-scale light source for an electron beam excitation-assisted (EXA) optical microscope. Three types of metals, Ag, Al, and Au, were compared, and an 181-fold enhancement of CL emission was achieved with Ag nanoparticles (NPs), with the plasmon resonance wavelength close to the emission wavelength energy of ZnO. The enhanced emission is plausibly attributed to LSP/exciton coupling. However, it is also attributed to an increase in coupling efficiency with penetration depth and also to an increase in light extraction efficiency by grading the refractive indices at the heterostructure.

The spatial coherence and the axial brightness of a cold field emission gun, a Schottky field emission gun and a $mathrm{LaB}_6$ thermionic gun are precisely measured. By analyzing the Airy pattern from a selected area aperture, various parameters including the spatial coherence length are determined. Using the determined coherence length, the axial brightness of the field emission guns is estimated using the equation which we previously derived based on the discussion of the Wigner function of an electron beam. We also make some extensions in the method to be applicable to the measurements of the thermionic gun, which has anisotropic intensity distribution in most case unlike the field emission guns. Not conventional average brightness but the axial brightness measured for the three kinds of emitters are compared accurately and precisely without influenced by the measurement conditions.