Sub-nanometer Spatial Resolution Research Articles

Multifunctional nanorobot system in precise evaluation and manipulation of virus capsid

It is imperative to have high adaptive techniques for sensing and manipulating biological targets at the nanoscale. This necessity becomes particularly crucial when dealing with fragile living bio-organisms like viruses, where the expression of capsids is closely linked to viral functions and genome constitution. Therefore, the development of a comprehensive system for dissecting and measuring viruses holds significant implications for the pharmaceutical industry and drug manufacturing. Leveraging the sub-nanometer spatial resolution and controllable tip-cantilever architecture of atomic force microscopy (AFM), a probe-laser system has been integrated as a self-sensing robotic end effector. To address intrinsic challenges in AFM-based robotic systems such as the lack of real-time monitoring, low scanning rates, and nonlinear motion caused by piezoelectric actuators, an augmented reality robotic system has been implemented. This system incorporates stereoscopic vision, a haptic feedback controller, a position recovery scheme, and a real-time force control algorithm. The integration of these components enhances the system’s capability to accurately dissect virus capsids. Operators can now perform highly efficient nanoscale tasks with multidimensional perception, utilizing the combination of stereoscopic vision and haptic force control. The position correction during manipulation can achieve a frame rate of over 30 frames per second, imperceptible to the operator, enabling closed-loop operation control. By adopting the proposed nanorobotic system in virology studies, it becomes possible to achieve accurate manipulation and dissection of SARS-CoV-2 pseudovirus capsids, and derive multi-parametric properties such as structural integrity, protein fragment thickness, and adhesive forces. The established nanobot system and experimental results serve as a guiding platform for high-accuracy evaluation in drug manufacturing development.

An Introduction to Magnetic Tweezers.

Magnetic tweezers are a single-molecule force and torque spectroscopy technique that enable the mechanical interrogation in vitro of biomolecules, such as nucleic acids and proteins. They use a magnetic field originating from either permanent magnets or electromagnets to attract a magnetic particle, thus stretching the tethering biomolecule. They nicely complement other force spectroscopy techniques such as optical tweezers and atomic force microscopy (AFM) as they operate as a very stable force clamp, enabling long-duration experiments over a very broad range of forces spanning from 10fN to 1nN, with 1-10milliseconds time and sub-nanometer spatial resolution. Their simplicity, robustness, and versatility have made magnetic tweezers a key technique within the field of single-molecule biophysics, being broadly applied to study the mechanical properties of, e.g., nucleic acids, genome processing molecular motors, protein folding,and nucleoprotein filaments. Furthermore, magnetic tweezers allow for high-throughput single-molecule measurements by tracking hundreds of biomolecules simultaneously both in real-time and at high spatiotemporal resolution. Magnetic tweezers naturally combine with surface-based fluorescence spectroscopy techniques, such as total internal reflection fluorescence microscopy, enabling correlative fluorescence and force/torque spectroscopy on biomolecules. This chapter presents an introduction to magnetic tweezers including a description of the hardware, the theory behind force calibration, its spatiotemporal resolution, combining it with other techniques, and a (non-exhaustive) overview of biological applications.

Electron Beam Induced Enhancement and Suppression of Oxidation in Cu Nanoparticles in Environmental Scanning Transmission Electron Microscopy.

We have investigated the effects of high-energy electron irradiation on the oxidation of copper nanoparticles in environmental scanning transmission electron microscopy (ESTEM). The hemispherically shaped particles were oxidized in 3 mbar of O2 in a temperature range 100-200 °C. The evolution of the particles was recorded with sub-nanometer spatial resolution in situ in ESTEM. The oxidation encompasses the formation of outer and inner oxide shells on the nanoparticles, arising from the concurrent diffusion of copper and oxygen out of and into the nanoparticles, respectively. Our results reveal that the electron beam actively influences the reaction and overall accelerates the oxidation of the nanoparticles when compared to particles oxidized without exposure to the electron beam. However, the extent of this electron beam-assisted acceleration of oxidation diminishes at higher temperatures. Moreover, we observe that while oxidation through the outward diffusion of Cu+ cations is enhanced, the electron beam appears to hinder oxidation through the inward diffusion of O2- anions. Our results suggest that the impact of the high-energy electrons in ESTEM oxidation of Cu nanoparticles is mostly related to kinetic energy transfer, charging, and ionization of the gas environment, and the beam can both enhance and suppress reaction rates.

First-Principles Simulations of Tip Enhanced Raman Scattering Reveal Active Role of Substrate on High-Resolution Images.

Tip-enhanced Raman scattering (TERS) has emerged as a powerful tool to obtain subnanometer spatial resolution fingerprints of atomic motion. Theoretical calculations that can simulate the Raman scattering process and provide an unambiguous interpretation of TERS images often rely on crude approximations of the local electric field. In this work, we present a novel and first-principles-based method to compute TERS images by combining Time Dependent Density Functional Theory (TD-DFT) and Density Functional Perturbation Theory (DFPT) to calculate Raman cross sections with realistic local fields. We present TERS results on free-standing benzene and C60 molecules, and on the TCNE molecule adsorbed on Ag(100). We demonstrate that chemical effects on chemisorbed molecules, often ignored in TERS simulations of larger systems, dramatically change the TERS images. This observation calls for the inclusion of chemical effects for predictive theory-experiment comparisons and an understanding of molecular motion at the nanoscale.

Correlating laser energy with compositional and atomic-level information of oxides in atom probe tomography

Atom probe tomography (APT) is a 3D analysis technique that offers unique chemical accuracy and sensitivity with sub-nanometer spatial resolution. There is an increasing interest in the application of APT to complex oxides materials, giving new insight into the relation between local variations in chemical composition and emergent physical properties. However, in contrast to the field of metallurgy, where APT is routinely applied to study materials at the atomic level, complex oxides and their specific field evaporation mechanisms are much less explored. Here, we perform APT measurements on the hexagonal manganite ErMnO3 and systematically study the effect of different experimental parameters on the measured composition and structure. We demonstrate that both the mass resolving power (MRP) and compositional accuracy can be improved by increasing the charge-state ratio (CSR) working at low laser energy (< 5 pJ) for a given fixed detection rate. Furthermore, we observe a substantial preferential retention of Er atoms, which is suppressed at higher CSRs. We explain our findings based on fundamental field evaporation concepts, expanding the knowledge about the impact of key experimental parameters and the field evaporation process in complex oxides in general.

The Landscape of Gold Nanocrystal Surface Chemistry.

ConspectusGold nanoparticles (AuNPs) exhibit unique size- and shape-dependent properties not obtainable at the macroscale. Gold nanorods (AuNRs), with their morphology-dependent optical properties, ability to convert light to heat, and high surface-to-volume ratios, are of great interest for biosensing, medicine, and catalysis. While the gold core provides many fascinating properties, this Account focuses on AuNP soft surface coatings, which govern the interactions of nanoparticles with the local environments. Postmodification of AuNP surface chemistry can greatly alter NP colloidal stability, nano-bio interactions, and functionality. Polyelectrolyte coatings provide controllable surface-coating thickness and charge, which impact the composition of the acquired corona in biological settings. Covalent modification, in which covalently bound ligands replace the original capping layer, is often performed with thiols and disulfides due to their ability to replace native coatings. N-heterocyclic carbenes and looped peptides expand the possible functionalities of the ligand layer.The characterization of surface ligands bound to AuNPs, in terms of ligand density and dynamics, remains a challenge. Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool for understanding molecular structures and dynamics. Our recent NMR work on AuNPs demonstrated that NMR data were obtainable for ligands on NPs with diameters up to 25 nm for the first time. This was facilitated by the strong proton NMR signals of the trimethylammonium headgroup, which are present in a distinct regime from other ligand protons' signals. Ligand density analyses showed that the smallest AuNPs (below 4 nm) had the largest ligand densities, yet spin-spin T2 measurements revealed that these smallest NPs also had the most mobile ligand headgroups. Molecular dynamics simulations were able to reconcile these seemingly contradictory results.While NMR spectroscopy provides ligand information averaged over many NPs, the ligand distribution on individual particles' surfaces must also be probed to fully understand the surface coating. Taking advantage of improvements in electron energy loss spectroscopy (EELS) detectors employed with scanning transmission electron microscopy (STEM), a single-layer graphene substrate was used to calibrate the carbon K-edge EELS signal, allowing quantitative imaging of the carbon atom densities on AuNRs with sub-nanometer spatial resolution. In collaboration with others, we revealed that the mean value for surfactant-bilayer-coated AuNRs had 10-30% reduced ligand density at the ends of the rods compared to the sides, confirming prior indirect evidence for spatially distinct ligand densities.Recent work has found that surface ligands on nanoparticles can, somewhat surprisingly, enhance the selectivity and efficiency of the electrocatalytic reduction of CO2 by controlling access to the active site, tuning its electronic and chemical environment, or denying entry to impurities that poison the nanoparticle surface to facilitate reduction. Looking to the future, while NMR and EELS are powerful and complementary techniques for investigating surface coatings on AuNPs, the frontier of this field includes the development of methods to probe the surface ligands of individual NPs in a high-throughput manner, to monitor nano-bio interactions within complex matrices, and to study structure-property relationships of AuNPs in biological systems.

A single molecule structural biology view of a transporter cycle.

As atomic structures of membrane proteins are readily resolved and predicted due to the recent advances in biophysical methods like cryo-EM as well as computational tools like AlphaFold, there emerges an urgent need to investigate the real-time structural dynamics at the single molecule level to obtain novel insights in the membrane structural biology. Here, we present a unique sample preparation method for high-speed atomic force microscopy (HS-AFM), which enabled us to immobilize membrane proteins in an extended lipid bilayer, while preserving their dynamics. This strategy bypasses the previous requirement of the dense packing of molecules. The approach allowed to acquire HS-AFM movies of individual transporters with increased resolution and in a physiologically more relevant setting. Using GltPh, a trimeric aspartate transporter, as a model protein, we demonstrate that HS-AFM combined with the presented strategy was able to track structural dynamics of individual protomers at sub-second temporal resolution and sub-nanometer spatial resolution for tens of seconds. Recent cryo-EM studies of GltPh revealed two major states of the protein, outward-facing (OF) and inward-facing (IF) states, as well as several conformational sub-states. Real-time transitions between these states were observed in the HS-AFM data with unprecedented detail. For data analysis, we developed a principal component analysis (PCA) based method to sort protomers at each time point into different conformations according to their structural features, reconstructing a structure-time trace for each protomer of a single molecule. Using the previously developed localization AFM (LAFM) method, we calculate Angstrom-resolution GltPh surface structures of single molecules as it transits through several conformational states in the transport cycle. The presented method enables direct observation and analysis of the structure and dynamics of individual unlabeled membrane proteins, unlocking the potential of HS-AFM in the study of single molecule structural biology.

Microfluidics-Based Force Spectroscopy Enables High-Throughput Force Experiments with Sub-Nanometer Resolution and Sub-Piconewton Sensitivity.

Several techniques have been established to quantify the mechanicals of single molecules. However, most of them show only limited capabilities of parallelizing the measurement by performing many individual measurements simultaneously. Herein, a microfluidics-based single-molecule force spectroscopy method, which achieves sub-nanometer spatial resolution and sub-piconewton sensitivity and is capable of simultaneously quantifying hundreds of single-molecule targets in parallel, is presented. It relies on a combination of total internal reflection microscopy and microfluidics, in which monodisperse fluorescent beads are immobilized on the bottom of a microfluidic channel by macromolecular linkers. Application of a flow generates a well-defined shear force acting on the beads, whereas the nanomechanical linker response is quantified based on the force-induced displacement of individual beads. To handle the high amount of data generated, a cluster analysis which is capable of a semi-automatic identification of measurement artifacts and molecular populations is implemented. The method is validated by probing the mechanical response polyethylene glycol linkers and binding strength of biotin-NeutrAvidin complexes. Two energy barriers (at 3 and 5.7 Å, respectively) in the biotin-NeutrAvidin interaction are resolved and the unfolding behavior of talin's rod domain R3 in the force range between 1 to ≈10 pN is probed.

Probing Local Ion Insertion through Strain-Current Correlation

Among different electrochemical energy storage devices, the ones that store a higher amount of energy (e.g., batteries) often undergo larger ion insertion-induced structural transformation and volume change, which results in lower rate performance. The ones that store a lower amount of energy (e.g., supercapacitors) usually show gradual structural change and smaller volume change. There is a strong correlation between electrochemistry and mechanical properties. Understanding the electro-chemo-mechanical coupling and controlling the ion insertion-induced strain is crucial for achieving high power and high energy storage devices. In-situ atomic force microscopy (AFM) is well suited to tackle this task as it allows tracking local volume changes and other mechanical responses sub-nanometer spatial resolution under the conditions close to the device operationIn this work, we monitored electrode volume change via in-situ AFM and demonstrated the electro-chemo-mechanical coupling behaviors during proton insertion into WO3 materials. The concept of mechanical cyclic voltammetry (mCV) curves was developed, and the relationship between electrochemical current and strain was investigated with simplified models. The results revealed multiple ion-intercalation processes with different mechanical responses are involved during electrode cycling. Local heterogeneity was investigated via mCV mapping, confirming that the charging mechanisms varied across the electrode. These local variations could be further correlated to local morphology, crystal orientations, or chemical compositions. We further demonstrate that the mCV approach is applicable to a variety of energy storage materials (e.g., birnessite and MXene) with the increasing complexity of current-deformation relationships.The work was supported by the Fluid Interface Reactions, Structures, and Transport (FIRST), an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences. Measurements were performed at the Center for Nanophase Materials Sciences (CNMS), which is sponsored at Oak Ridge National Laboratory by the Scientific User Facilities Division, Office of Basic Energy Sciences.

Electron Microscopy Probing Electron-Photon Interactions in SiC Nanowires with Ultrawide Energy and Momentum Match.

Light-matter interactions are commonly probed by optical spectroscopy, which, however, has some fundamental limitations such as diffraction-limited spatial resolution, tiny momentum transfer, and noncontinuous excitation/detection. In this work, through the use of scanning transmission electron microscopy-electron energy loss spectroscopy (STEM-EELS) with ultrawide energy and momentum match and subnanometer spatial resolution, the longitudinal Fabry-Perot (FP) resonating modes and the transverse whispering-gallery modes (WGMs) in individual SiC nanowires are simultaneously excited and detected, which span from near-infrared (∼1.2 μm) to ultraviolet (∼0.2 μm) spectral regime, and the momentum transfer can range up to 108 cm-1. The size effects on the resonant spectra of nanowires are also revealed. This work provides an alternative technique to optical resonating spectroscopy and light-matter interactions in dielectric nanostructures, which is promising for modulating free electrons via photonic structures.

Sub-Nanometer Resolved Tip-Enhanced Raman Spectroscopy of a Single Molecule on the Si(111) Substrate

The investigation of molecular adsorption structures on nonmetallic surfaces is important for understanding the functionalities of nanodevices and biomolecules. In contrast to extensive tip-enhanced Raman spectroscopy (TERS) studies on metallic substrates with strong plasmonic enhancement, the studies on nonmetallic substrates like silicon (Si) are relatively limited. It remains to be clarified whether it is possible to detect the TERS signal from a single molecule directly adsorbed on a nonmetallic surface and how precise its spatial resolution could be achieved. Here we use Si(111) as a prototype nonmetallic substrate without plasmonic responses (in the visible region) and investigate the TERS of single bis(phthalocyaninato) terbium(III) (TbPc2) molecules adsorbed on this surface. We can not only detect the TERS signal from a single TbPc2 molecule, but also specify distinct adsorption configurations according to different TERS spectral features. The spatial resolution of TERS is also found down to ∼0.6 nm, which is slightly poorer than but still comparable with the resolution for the TbPc2 molecule directly adsorbed on the silver substrate. By combining with theoretical simulations, we find that although a plasmonic substrate can induce a larger TERS enhancement than a dielectric substrate, the subnanometer spatial resolution of TERS is mainly determined by the atomistic protrusion at the apex of a plasmonic metal tip, regardless of whether the substrate is plasmonic or dielectric. These findings offer a promise for applying subnanometer resolved TERS to a wide range of systems that contain nonmetallic substrates, such as in the semiconductor industry, 2D materials, and biomedical science.

Sub-nanometer mapping of strain-induced band structure variations in planar nanowire core-shell heterostructures

Strain relaxation mechanisms during epitaxial growth of core-shell nanostructures play a key role in determining their morphologies, crystal structure and properties. To unveil those mechanisms, we perform atomic-scale aberration-corrected scanning transmission electron microscopy studies on planar core-shell ZnSe@ZnTe nanowires on α-Al2O3 substrates. The core morphology affects the shell structure involving plane bending and the formation of low-angle polar boundaries. The origin of this phenomenon and its consequences on the electronic band structure are discussed. We further use monochromated valence electron energy-loss spectroscopy to obtain spatially resolved band-gap maps of the heterostructure with sub-nanometer spatial resolution. A decrease in band-gap energy at highly strained core-shell interfacial regions is found, along with a switch from direct to indirect band-gap. These findings represent an advance in the sub-nanometer-scale understanding of the interplay between structure and electronic properties associated with highly mismatched semiconductor heterostructures, especially with those related to the planar growth of heterostructured nanowire networks.

(Invited) Development of THz-Field-Driven Scanning Tunneling Luminescence Spectroscopy for Future Investigation of Exciton Dynamics

Exciton dynamics is responsible for the photophysical and photochemical functions, such as energy harvesting, light emission, and photosynthesis. In particular, energy conversions, transfers, and dissipations at interfaces and nanostructures govern the dynamics of excitons. Nevertheless, the spatial resolutions of conventional optical methods are limited to several hundred nanometers due to the diffraction limit of light. Luminescence measurements combined with scanning tunneling microscopy (STM) enable to investigate optical properties with sub-nanometer spatial resolution in various systems such as metals[1], semiconductors[2], 2D materials[3], and molecules[4-5]. In this method, the scanning tunneling luminescence (STL) spectroscopy, photons induced by the tunneling current of STM are detected. Since the tunneling current can be controlled with atomic spatial resolution, it is possible to investigate optical phenomena in localized regions. While STL spectroscopy is a powerful tool for investigating exciton dynamics with atomic-scale spatial resolution, the time resolution of STL spectroscopy is limited because it uses “steady-state” tunneling current for the excitation. Therefore, this method cannot be applicable to track the ultrafast exciton dynamics in real-time.For overcoming the limitation of time resolution of STM, a single-cycle terahertz (THz) electric field has recently been installed to drive tunneling electrons[6-9]. By coupling the THz pulse to an atomically sharp metal tip of STM, the electric field is confined to the tip, and it enables to manipulate the tunneling electrons with extremely high spatiotemporal resolution. This method, THz-STM, has been utilized to track the ultrafast carrier dynamics[6-8] and motion of a molecule[9] with sub-picosecond time resolution. In this work, we combined a photon detection system with THz-STM to establish THz-STL spectroscopy (Fig. 1a). We measured visible photon emission from the radiative decay of a localized plasmon as the first demonstration of THz-STL spectroscopy[10]. We anticipate that the THz-STL spectroscopy will open new avenues for “real-time” and “real-space” investigations of exciton dynamics in near future.In this work, we generated single-cycle THz pulses via optical rectification of laser pulses from a Yb fiber laser in a LiNbO3 prism using a tilted-pulse-front configuration (Fig. 1b). The generated THz pulses were guided into a low-temperature STM and focused to the STM junction (Fig. 1a). Figure 1c shows a current trace on Ag(111) surface, where the current induced by the THz pulses was measured as 2.65 pA. Based on the simulation of electron tunneling probability, the maximum magnitude of the voltage applied by the THz pulse was estimated to be ~6.5 V. Figure 1d shows STL spectra when the THz pulse was either introduced to the STM (red) or blocked (grey). Whereas no peak is seen in the grey spectrum, a broad peak ranging from 1.3 eV to 2.3 eV appears in the red spectrum, which is originated from the radiative decay of a plasmon localized in the gap.In this presentation, we would like to discuss the detailed mechanism of plasmon excitation by THz-field-driven electrons as well as the challenges of applying THz-STL to the real-time investigation of exciton dynamics in a molecule.[1] R. Berndt et al., Phys. Rev. Lett. 67, 3796 (1991). [2] J. K. Gimzewski et al., Z. Phys. B Condensed Matter 72, 497 (1988). [3] Schuler et al., Sci. Adv. 6, eabb5988 (2020). [4] H. Imada et al., Nature 538, 364 (2016). [5] K. Kimura et al., Nature 570, 210 (2019). [6] T. L. Cocker et al., Nat. Photon. 7, 620 (2013). [7] K. Yoshioka et al., Nat. Photon. 10, 762 (2016). [8] V. Jelic et al., Nature Physics 13, 591 (2017). [9] D. Peller et al., Nature 585, 58 (2020). [10] K. Kimura et al., ACS photon. 8, 982 (2021). Figure 1

Nanoscale-Resolved Surface-to-Bulk Electron Transport in CsPbBr3 Perovskite.

Describing the nanoscale charge carrier transport at surfaces and interfaces is fundamental for designing high-performance optoelectronic devices. To achieve this, we employ time- and angle-resolved photoelectron spectroscopy with ultraviolet pump and extreme ultraviolet probe pulses. The resulting high surface sensitivity reveals an ultrafast carrier population decay associated with surface-to-bulk transport, which was tracked with a sub-nanometer spatial resolution normal to the surface, and on a femtosecond time scale, in the case of the inorganic CsPbBr3 lead halide perovskite. The decay time exhibits a pronounced carrier density dependence, which is attributed via modeling to enhanced diffusive transport and concurrent recombination. The transport is found to approach an ordinary diffusive regime, limited by electron–hole scattering, at the highest excitation fluences. This approach constitutes an important milestone in our capability to probe hot-carrier transport at solid interfaces with sub-nanometer resolution in a theoretically and experimentally challenging, yet technologically relevant, high-carrier-density regime.

Improving Quantitative EDS Chemical Analysis of Alloy Nanoparticles by PCA Denoising: Part I, Reducing Reconstruction Bias

Scanning transmission electron microscopy is a crucial tool for nanoscience, achieving sub-nanometric spatial resolution in both image and spectroscopic studies. This generates large datasets that cannot be analyzed without computational assistance. The so-called machine learning procedures can exploit redundancies and find hidden correlations. Principal component analysis (PCA) is the most popular approach to denoise data by reducing data dimensionality and extracting meaningful information; however, there are many open questions on the accuracy of reconstructions. We have used experiments and simulations to analyze the effect of PCA on quantitative chemical analysis of binary alloy (AuAg) nanoparticles using energy-dispersive X-ray spectroscopy. Our results demonstrate that it is possible to obtain very good fidelity of chemical composition distribution when the signal-to-noise ratio exceeds a certain minimal level. Accurate denoising derives from a complex interplay between redundancy (data matrix size), counting noise, and noiseless data intensity variance (associated with sample chemical composition dispersion). We have suggested several quantitative bias estimators and noise evaluation procedures to help in the analysis and design of experiments. This work demonstrates the high potential of PCA denoising, but it also highlights the limitations and pitfalls that need to be avoided to minimize artifacts and perform reliable quantification.

Competing oxidation mechanisms in Cu nanoparticles and their plasmonic signatures.

Chemical reactions involving nanoparticles often follow complex processes. In this respect, real-time probing of single nanoparticles under reactive conditions is crucial for uncovering the mechanisms driving the reaction pathway. Here, we have captured in situ the oxidation of single Cu nanoparticles to unravel a sequential competitive activation of different mechanisms at temperatures 50-200 °C. Using environmental scanning transmission electron microscopy, we monitor the evolution of oxide formation with sub-nanometre spatial resolution, and show how the prevalence of oxide island nucleation, Cabrera-Mott, Valensi-Carter and Kirkendall mechanisms under different conditions determines the morphology of the particles. Moreover, using in situ electron energy-loss spectroscopy, we probe the localised surface plasmons of individual particles during oxidation, and with the aid of finite-difference time-domain electrodynamic simulations investigate the signature of each mechanism in their plasmonic response. Our results shed light on the rich and intricate processes involved in the oxidation of nanoparticles, and provide in-depth insight into how these processes govern their morphology and optical response, beneficial for applications in catalysis, sensing, nanomedicine and plasmonics.

Robust autofocusing for scanning electron microscopy based on a dual deep learning network

Scanning electron microscopy (SEM) is a high-resolution imaging technique with subnanometer spatial resolution that is widely used in materials science, basic science, and nanofabrication. However, conducting SEM is rather complex due to the nature of using an electron beam and the many parameters that must be adjusted to acquire high-quality images. Only trained operators can use SEM equipment properly, meaning that the use of SEM is restricted. To broaden the usability of SEM, we propose an autofocus method for a SEM system based on a dual deep learning network, which consists of an autofocusing-evaluation network (AENet) and an autofocusing-control network (ACNet). The AENet was designed to evaluate the quality of given images, with scores ranging from 0 to 9 regardless of the magnification. The ACNet can delicately control the focus of SEM online based on the AENet’s outputs for any lateral sample position and magnification. The results of these dual networks showed successful autofocus performance on three trained samples. Moreover, the robustness of the proposed method was demonstrated by autofocusing on unseen samples. We expect that our autofocusing system will not only contribute to expanding the versatility of SEM but will also be applicable to various microscopes.

Probing Local Ion Insertion via Electro-Chemo-Mechanical Coupling Responses

Electrochemical energy storage devices (e.g., Li-ion batteries and supercapacitors) store energy through electrolytic ion insertion in the electrode. During the ion insertion process, the electrode needs to open up or reorganize its crystallographic structure in order to host the ions, which results in macroscopic and microscopic deformation. The amount of energy barrier required for electrode structural change during the ion insertion process dictates the energy and power density and cycle life of the device. Therefore, understanding the electrode deformation and changes in mechanical responses during electrochemical events and their impact on devices’ performance is crucial for achieving high power and high energy storage devices. In-situ atomic force microscopy (AFM) is well suited to tackle this task as it allows tracking local volume changes and other mechanical responses sub-nanometer spatial resolution under the conditions close to the device operationIn this work, we demonstrate the electro-chemo-mechanical coupling behaviors of proton insertion into WO3 materials via in-situ AFM. The concept of mechanical cyclic voltammetry (mCV) curves was developed, and the relationship between electrochemical current and strain were investigated with simplified models. The results revealed multiple ion-intercalation process with different mechanical responses are involved during electrode cycling. Local heterogeneity was investigated via mCV mapping, confirming that the charging mechanisms varied across the electrode. These local variations could be further corelated to local morphology, crystal orientations or chemical compositions. Besides providing information of redox heterogeneity, the mechanical CV methodology allows us to circumvent the common issues that are often encountered in the global electrochemical characterizations (ex: cell resistance and unwanted parasitic reaction). We further demonstrate that the mCV approach is applicable to a variety of energy storage materials with increasing complexity of current-deformation relationships.The work was supported by the Fluid Interface Reactions, Structures and Transport (FIRST), an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences. Measurements were performed at the Center for Nanophase Materials Sciences (CNMS), which is sponsored at Oak Ridge National Laboratory by the Scientific User Facilities Division, Office of Basic Energy Sciences.

Perovskite Quantum Dots for Super‐Resolution Optical Microscopy: Where Strong Photoluminescence Blinking Matters (Advanced Optical Materials 18/2021)

Direct stochastic optical reconstruction microscopy can super-resolve the location of blinking nanoscale emitters with spatial resolutions of a few tens of nanometers. As bright and photostable emitters, colloidal perovskite quantum dots are a promising alternative to organic molecules, but they suffer from nonoptimal blinking statistics. In article number 2100620, Gabriele Rainò, Maksym V. Kovalenko and co-workers outline how blinking-engineered perovskite quantum dots can be employed in stochastic optical reconstruction microscopy towards sub-nanometer spatial resolution.

Studying the rhodopsin-like G protein-coupled receptors by atomic force microscopy.

Rhodopsin-like G protein-coupled receptors (GPCRs), widely distributed in microorganisms, invertebrates, and vertebrates, are the largest class in GPCRs, and are involved in many important physiological and pathological processes, including the photosensitivity, regulation of behavior and emotion, and so on. Atomic force microscopy (AFM) is a powerful and multifunctional toolkit in bionanotechnology, as it can image the morphology of membrane proteins at subnanometer spatial resolution and detect forces related with membrane proteins down to piconewton level by single-molecule force spectroscopy (SMFS) mode under physiological conditions. Herein, the achievements of AFM in the study of rhodopsin-like GPCRs, including observing the high-resolution topography and structural changes, revealing the interaction forces, binding kinetics, and mechanical properties (such as modulus), are reviewed and summarized. Finally, the challenges, outlook, and prospects of AFM in the study of rhodopsin-like GPCRs are discussed.