The crystallographic analysis of nanoscale phases with dimensions well below the spatial probing volume of electron backscatter diffraction (EBSD) traditionally rely on electron microscopy in transmission (either in SEM or TEM), because EBSD patterns are invariably dominated by the matrix phase contribution and present seemingly no trace from such nanoscale phases. Yet, this study shows that such nanoscale features generate a very faint but valuable secondary diffraction signal which can be retrieved. A diffraction pattern postprocessing method is presented which focuses on the detection of such secondary signal emitted by nanoscale minority phases in overlapped patterns dominated by a dominant matrix signal. The predominant, majority phase contribution in EBSD patterns is removed by a close-neighbor pattern subtraction routine, after which both the conventional Hough indexing method as well as pattern matching methods can be used to reveal the crystallography, spatial distribution, morphology, and orientation of nanoscale minority phases initially absent from EBSD maps. Nanolamellar pearlitic steel, which has long been out of reach for EBSD, has been chosen as an application example.

As a real-space technique, atomic-resolution STEM imaging contains both amplitude and geometric phase information about structural order in materials, with the latter encoding important information about local variations and heterogeneities present in crystalline lattices. Such phase information can be extracted using geometric phase analysis (GPA), a method which has generally focused on spatially mapping elastic strain. Here we demonstrate an alternative phase demodulation technique and its application to reveal complex structural phenomena in correlated quantum materials. As with other methods of image phase analysis, the phase lock-in approach can be implemented to extract detailed information about structural order and disorder, including dislocations and compound defects in crystals. Extending the application of this phase analysis to Fourier components that encode periodic modulations of the crystalline lattice, such as superlattice or secondary frequency peaks, we extract the behavior of multiple distinct order parameters within the same image, yielding insights into not only the crystalline heterogeneity but also subtle emergent order parameters such as antipolar displacements. When applied to atomic-resolution images spanning large (~0.5 × 0.5 μm2) fields of view, this approach enables vivid visualizations of the spatial interplay between various structural orders in novel materials.

A high-frequency short-pulsed stroboscopic micro-visual system was employed to capture the transient image sequences of a periodically in-plane working micro-electro-mechanical system (MEMS) devices. To demodulate the motion parameters of the devices from the images, we developed the feature point matching (FPM) algorithm based on Speeded-Up Robust Features (SURF). A MEMS gyroscope, vibrating at a frequency of 8.189 kHz, was used as a testing sample to evaluate the performance of the proposed algorithm. Within the same processing time, the SURF-based FPM method demodulated the velocity of the in-plane motion with a precision of 10−5 pixels of the image, which was two orders of magnitude higher than the template-matching and frame-difference algorithms.

Using a combination of simulated data and pyrite isotopic reference materials, we have refined a methodology to obtain quantitative δ34S measurements from atom probe tomography (APT) datasets. This study builds on previous attempts to characterize relative 34S/32S ratios in gold-containing pyrite using APT. We have also improved our understanding of the artifacts inherent in laser-pulsed APT of insulators. Specifically, we find the probability of multi-hit detection events increases during the APT experiment, which can have a detrimental effect on the accuracy of the analysis. We demonstrate the use of standardized corrected time-of-flight single-hit data for our isotopic analysis. Additionally, we identify issues with the standard methods of extracting background-corrected counts from APT mass spectra. These lead to inaccurate and inconsistent isotopic analyses due to human variability in peak ranging and issues with background correction algorithms. In this study, we use the corrected time-of-flight single-hit data, an adaptive peak fitting algorithm, and an improved deconvolution algorithm to extract 34S/32S ratios from the S2+ peaks. By analyzing against a standard material, acquired under similar conditions, we have extracted δ34S values to within ±5‰ (1‰ = 1 part per thousand) of the published values of our standards.

Analytical studies of nanoparticles (NPs) are frequently based on huge datasets derived from hyperspectral images acquired using scanning transmission electron microscopy. These large datasets require machine learning computational tools to reduce dimensionality and extract relevant information. Principal component analysis (PCA) is a commonly used procedure to reconstruct information and generate a denoised dataset; however, several open questions remain regarding the accuracy and precision of reconstructions. Here, we use experiments and simulations to test the effect of PCA processing on data obtained from AuAg alloy NPs a few nanometers wide with different compositions. This study aims to address the reliability of chemical quantification after PCA processing. Our results show that the PCA treatment mitigates the contribution of Poisson noise and leads to better quantification, indicating that denoised results may be reliable from the point of view of both uncertainty and accuracy for properly planned experiments. However, the initial data need to be of sufficient quality: these results can only be obtained if the signal-to-noise ratio of input data exceeds a minimal value to avoid the occurrence of random noise bias in the PCA reconstructions.

The Compton signal in electron energy loss spectroscopy (EELS) is used to determine the projected electron momentum density of states for the solid. A frequent limitation however is the strong dynamical scattering of the incident electron beam within a crystalline specimen, i.e. Bragg diffracted beams can be additional sources of Compton scattering that distort the measured profile from its true shape. The Compton profile is simulated via a multislice method that models dynamical scattering both before and after the Compton energy loss event. Simulations indicate the importance of both the specimen illumination condition and EELS detection geometry. Based on this, a strategy to minimize diffraction artifacts is proposed and verified experimentally. Furthermore, an inversion algorithm to extract the projected momentum density of states from a Compton measurement performed under strong diffraction conditions is demonstrated. The findings enable a new route to more accurate electron Compton data from crystalline specimens.

The surging interest in manipulating the polarization of piezo/ferroelectric materials by means of light has driven an increasing number of studies toward their light-polarization interaction. One way to investigate such interaction is by performing piezoresponse force microscopy (PFM) while/after the sample is exposed to light illumination. However, caution must be exercised when analyzing and interpreting the data, as demonstrated in this paper, because sizeable photo-response observed in the PFM amplitude image of the sample is shown to be caused by the electrostatic interaction between the photo-induced surface charge and tip. Through photo-assisted Kelvin probe force microscopy (KPFM), positive surface potential is found to be developed near the sample's surface under 405 nm light illumination, whose effects on the measured PFM signal is revealed by the comparative studies on its amplitude curves that are obtained using PFM spectroscopy mode with/without illumination. This work exemplifies the need for complementary use of KPFM, PFM imaging mode, and PFM spectroscopy mode in order to distinguish real behavior from artifacts.

Low-voltage scanning electron microscopes (LV-SEMs) are widely used in nanoscience. However, image resolution for SEMs is restricted by chromatic aberration due to energy spread of the electron beam at low acceleration voltage. This study introduces a new monochromator (MC) with offset cylindrical lenses (CLs) as one solution for LV-SEMs. The MC optics, with highly excited CLs in offset layouts, has advantageous high performance and simple experimental setup, making it suitable for field emission LV-SEMs. In a preliminary evaluation, our MC reduced the energy spread from 770 to 67 meV. The MC was integrated into a commercial SEM equipped with an out-lens (a conventional objective lens without immersion magnetic or retarding electric fields) and an Everhart–Thornley detector. Comparing SEM images under two conditions with the MC turned on or off, the spatial resolution was improved by 58% at 0.5 and 1 keV. The filtering effect of the MC decreased the probe current with a ratio (i.e., transmittance) of 5.7%, which was consistent with estimations based on measured energy spreads. To the best of our knowledge, this is the first report on an effective MC with higher-energy resolution than 100 meV and the results offer encouraging prospects for LV-SEM technology.

A structured-light projection system was designed for microscale objects with surface heights that ranged from tens to hundreds of microns. The system was composed of a universal projector and microscope system that supported editing the attributes of structured-light patterns in real-time and was capable of projecting microscale patterns. On this basis, reconstructing the metal surfaces of microscale objects based on grid patterns of structured light was investigated, the internal and external parameters of microscope vision and projection systems were calibrated, and an image algorithm for grid-node detection was designed. The results indicated that the proposed method successfully reconstructed the three-dimensional (3D) surface of microscale objects, and the reconstruction results were consistent with the original surfaces. With 95% confidence, the reconstruction precision in the X- and Y-directions was approximately ±4.0 μm and in the Z-direction was approximately ±7.5 μm. The designed system and the proposed method were suitable for 3D-shape measurement of microstructures in microscopic fields and can be adapted to meet a broader range of applications, as compared to current methods.

Reliable spatially resolved compositional analysis through atom probe tomography requires an accurate placement of the detected ions within the three-dimensional reconstruction. Unfortunately, for heterogeneous systems, traditional reconstruction protocols are prone to position some ions incorrectly. This stems from the use of simplified projection laws which treat the emitter apex as a spherical cap, although the actual shape may be far more complex. For instance, sampled materials with compositional heterogeneities are known to develop local variations in curvature across the emitter due to their material phase specific evaporation fields. This work provides three pivotal precursors to improve the spatial accuracy of the reconstructed volume in such cases. First, we show scanning probe microscopy enables the determination of the local curvature of heterogeneous emitters, thus providing the essential information for a more advanced reconstruction considering the actual shape. Second, we demonstrate the cyclability between scanning probe characterization and atom probe analysis. This is a key ingredient of more advanced reconstruction protocols whereby the characterization of the emitter topography is executed at multiple stages of the atom probe analysis. Third, we show advances in the development of an electrostatically driven reconstruction protocol which are expected to enable reconstruction based on experimental tip shapes.

Maintaining telecentricity and zooming in microscopic systems with prolonged depths of focus is a difficult challenge because these properties degrade while moving to different axial planes in the extended focal depth. In this paper, we propose the proof of concept for an automated dual-mode microscopic system that combines two electrically tunable lenses (ETLs) with a variable numerical aperture controller placed. It acts as a viable solution to allow both multiplane microscopic zooming and telecentricity with consistent image resolution throughout the objective's extended focal depth. The image plane remains fixed for both the modes of operation, namely telecentricity and multiplane zooming. To validate the performance of the proposed idea, both simulations and experiments are carried out at various ETL curvature ranges. Over the whole zoom distance range, the experimental zoom ratio is determined to range from −2.723X to −34.42X. The experimental and simulation findings are compared and found to be quite similar, with magnification error percentages of 2.26% for zoom mode and 1.27% for telecentric mode. The comprehensive explanation of simulation and experimental results demonstrate the feasibility of the proposed method for both multiplane zoom and telecentric operations on a single platform in microscopic applications.

Precision and accuracy of quantitative scanning transmission electron microscopy (STEM) methods such as ptychography, and the mapping of electric, magnetic, and strain fields depend on the dose. Reasonable acquisition time requires high beam current and the ability to quantitatively detect both large and minute changes in signal. A new hybrid pixel array detector (PAD), the second-generation Electron Microscope Pixel Array Detector (EMPAD-G2), addresses this challenge by advancing the technology of a previous generation PAD, the EMPAD. The EMPAD-G2 images continuously at a frame-rates up to 10 kHz with a dynamic range that spans from low-noise detection of single electrons to electron beam currents exceeding 180 pA per pixel, even at electron energies of 300 keV. The EMPAD-G2 enables rapid collection of high-quality STEM data that simultaneously contain full diffraction information from unsaturated bright-field disks to usable Kikuchi bands and higher-order Laue zones. Test results from 80 to 300 keV are presented, as are first experimental results demonstrating ptychographic reconstructions, strain and polarization maps. We introduce a new information metric, the maximum usable imaging speed (MUIS), to identify when a detector becomes electron-starved, saturated or its pixel count is mismatched with the beam current.

As the measurement scale shrinks, the reliability of nanoscale measurement is even more crucial for a variety of applications, including semiconductor electronics, optical metamaterials, and sensors. Specifically, it is difficult to measure the nanoscale morphology at the exact location though it is required for novel applications based on hybrid nanostructures combined with 2D materials. Here, we introduce an advanced hybrid positioning system to measure the region of interest with enhanced speed and high precision. A 5-axis positioning stage (XYZ, R, gripper) makes it possible to align the sample within a 10-μm field of view (FOV) in both the scanning electron microscope (SEM) and the atomic force microscope (AFM). The reproducibility of the sample position was investigated by comparing marker patterns and denting points between the SEM and AFM, revealing an accuracy of 6.5 ± 2.1 μm for the x-axis and 4.5 ± 1.7 μm for the y-axis after 12 repetitions. By applying a different measurement process according to the characteristics of 2D materials, various information such as height, length, or roughness about MoTe2 rods and MoS2 film was obtained in the same measurement area. As a consequence, overlaid two images can be obtained for detailed information about 2D materials.

Vaginitis is a prevalent gynecologic disease that threatens millions of women’s health. Although microscopic examination of vaginal discharge is an effective method to identify vaginal infections, manual analysis of microscopic leucorrhea images is extremely time-consuming and labor-intensive. To automate the detection and identification of visible components in microscopic leucorrhea images for early-stage diagnosis of vaginitis, we propose a novel end-to-end deep learning-based cells detection framework using attention-based detection with transformers (DETR) architecture. The transfer learning was applied to speed up the network convergence while maintaining the lowest annotation cost. To address the issue of detection performance degradation caused by class imbalance, the weighted sampler with on-the-fly data augmentation module was integrated into the detection pipeline. Additionally, the multi-head attention mechanism and the bipartite matching loss system of the DETR model perform well in identifying partially overlapping cells in real-time. With our proposed method, the pipeline achieved a mean average precision (mAP) of 86.00% and the average precision (AP) of epithelium, leukocyte, pyocyte, mildew, and erythrocyte was 96.76, 83.50, 74.20, 89.66, and 88.80%, respectively. The average test time for a microscopic leucorrhea image is approximately 72.3 ms. Currently, this cell detection method represents state-of-the-art performance.

Electron tomography (ET) has gained increasing attention for the 3D characterization of nanoparticles. However, the missing wedge problem due to a limited tilt angle range is still the main challenge for accurate reconstruction in most experimental TEM setups. Advanced algorithms could in-paint or compensate to some extent the missing wedge artifacts, but cannot recover the missing structural information completely. 360° ET provides an option to solve this problem by tilting a needle-shaped specimen over the full tilt range and thus filling the missing information. However, sample preparation especially for fine powders to perform full-range ET is still challenging, thus limiting its application. In this work, we propose a new universal sample preparation method that enables the transfer of selected individual nanoparticle or a few separated nanoparticles by cutting a piece of carbon film supporting the specimen particles and mounting them onto the full-range tomography holder tip with the help of an easily prepared sharp tungsten tip. This method is demonstrated by 360° ET of Pt@TiO2 hollow cage catalyst showing high quality reconstruction without missing wedge.

The objective of this work was to characterize the ontogenesis of Protium spruceanum secretory ducts, to evaluate the effects of seasonality on that process, and to characterize the chemical nature of the resin. Morphometric, anatomical, micromorphometric, histochemical, and ultrastructural evaluations of shoot apexes and chemical analyses of the resin were performed. The ducts of schizolysigenous origin are distributed in the primary and secondary phloem. The subsecretory tissue is meristematic and can restore the secretory epithelium. Secretory epithelial cells have wall thickening resembling that of the Casparian strip that regulates secretion reflux. The main resin compounds are pentacyclic triterpenoids, α- and β-amyrins, and α- and β-amyrenones, which are reported here for the first time for this species. The presence of electron-dense and electron-opaque structures, in the secretory epithelial cells, are compatible with the triterpenes and mucilage identified in the resin. Rising temperatures, rainfall, and increasing day length induce the formation of ducts in the vascular cambium throughout Spring/Summer. The abundant production of resin rich in pentacyclic triterpenes indicates the potential use of the species for medicinal and cosmetic purposes. The understanding that secretory processes are concentrated during the Spring/Summer seasons will contribute to the definition of resin extraction management strategies.

Imaging of liquids and cryogenic biological materials by electron microscopy has been recently enabled by innovative approaches for specimen preparation and the fast development of optimized instruments for cryo-enabled electron microscopy (cryo-EM). Yet, cryo-EM typically lacks advanced analytical capabilities, in particular for light elements. With the development of protocols for frozen wet specimen preparation, atom probe tomography (APT) could advantageously complement insights gained by cryo-EM. Here, we report on different approaches that have been recently proposed to enable the analysis of relatively large volumes of frozen liquids from either a flat substrate or the fractured surface of a wire. Both allowed for analyzing water ice layers which are several micrometers thick consisting of pure water, pure heavy water, and aqueous solutions. We discuss the merits of both approaches and prospects for further developments in this area. Preliminary results raise numerous questions, in part concerning the physics underpinning field evaporation. We discuss these aspects and lay out some of the challenges regarding the APT analysis of frozen liquids.