Liquid-phase Electron Microscopy Research Articles

Discovery of Molecular Intermediates and Nonclassical Nanoparticle Formation Mechanisms by Liquid Phase Electron Microscopy and Reaction Throughput Analysis

Formation kinetics of metal nanoparticles are generally described via mass transport and thermodynamics‐based models, such as diffusion‐limited growth and classical nucleation theory (CNT). However, metal monomers are commonly assumed as precursors, leaving the identity of molecular intermediates and their contribution to nanoparticle formation unclear. Herein, liquid phase transmission electron microscopy (LPTEM) and reaction kinetic modeling are utilized to establish the nucleation and growth mechanisms and discover molecular intermediates during silver nanoparticle formation. Quantitative LPTEM measurements show that their nucleation rate decreases while growth rate is nearly invariant with electron dose rate. Reaction kinetic simulations show that Ag4 and Ag− follow a statistically similar dose rate dependence as the experimentally determined growth rate. We show that experimental growth rates are consistent with diffusion‐limited growth via the attachment of these species to nanoparticles. The dose rate dependence of nucleation rate is inconsistent with CNT. A reaction‐limited nucleation mechanism is proposed and it is demonstrated that experimental nucleation kinetics are consistent with Ag42+ aggregation rates at millisecond time scales. Reaction throughput analysis of the kinetic simulations uncovered formation and decay pathways mediating intermediate concentrations. We demonstrate the power of quantitative LPTEM combined with kinetic modeling for establishing nanoparticle formation mechanisms and principal intermediates.

Discovering Nanoparticle Formation Mechanisms and Molecular Intermediates with Liquid Phase Electron Microscopy and Reaction Networks

Discovering Nanoparticle Formation Mechanisms and Molecular Intermediates with Liquid Phase Electron Microscopy and Reaction Networks

Nanopipette dynamic microscopy unveils nano coffee ring

Liquid-phase electron microscopy (LP-EM) imaging has revolutionized our understanding of nanosynthesis and assembly. However, the current closed geometry limits its application for open systems. The ubiquitous physical process of the coffee-ring phenomenon that underpins materials and engineering science remains elusive at the nanoscale due to the lack of experimental tools. We introduce a quartz nanopipette liquid cell with a tunable dimension that requires only standard microscopes. Depending on the imaging condition, the open geometry of the nanopipette allows the imaging of evaporation-induced pattern formation, but it can also function as an ordinary closed-geometry liquid cell where evaporation is negligible despite the nano opening. The nano coffee-ring phenomenon was observed by tracking individual nanoparticles in an evaporating nanodroplet created from a thin liquid film by interfacial instability. Nanoflows drive the assembly and disruption of a ring pattern with the absence of particle-particle correlations. With surface effects, nanoflows override thermal fluctuations at tens of nanometers, in which nanoparticles displayed a "drunken man trajectory" and performed work at a value much smaller than kBT.

Imaging biomacromolecules in action with liquid-phase electron microscopy

Imaging biomacromolecules in action in a liquid environment poses a challenge to single-molecule experiments. Liquid-phase electron microscopy (LP-EM) has demonstrated its unique ability to reveal previously unseen transient dynamics and intermediate states of delicate biomacromolecular systems (protein, DNA/RNA) with deep-learning methods and careful evaluation of electron beam effects.

Electron videography of a lipid-protein tango.

Biological phenomena, from enzymatic catalysis to synaptic transmission, originate in the structural transformations of biomolecules and biomolecular assemblies in liquid water. However, directly imaging these nanoscopic dynamics without probes or labels has been a fundamental methodological challenge. Here, we developed an approach for "electron videography"-combining liquid phase electron microscopy with molecular modeling-with which we filmed the nanoscale structural fluctuations of individual, suspended, and unlabeled membrane protein nanodiscs in liquid. Systematic comparisons with biochemical data and simulation indicate the graphene encapsulation involved can afford sufficiently reduced effects of the illuminating electron beam for these observations to yield quantitative fingerprints of nanoscale lipid-protein interactions. Our results suggest that lipid-protein interactions delineate dynamically modified membrane domains across unexpectedly long ranges. Moreover, they contribute to the molecular mechanics of the nanodisc as a whole in a manner specific to the protein within. Overall, this work illustrates an experimental approach to film, quantify, and understand biomolecular dynamics at the nanometer scale.

Revealing Catalyst Self-Adjustment in C–S Cross-Coupling through Multiscale Liquid-Phase Electron Microscopy

Revealing Catalyst Self-Adjustment in C–S Cross-Coupling through Multiscale Liquid-Phase Electron Microscopy

Graphene Electrode for Studying CO2 Electroreduction Nanocatalysts under Realistic Conditions in Microcells.

The ability to resolve the dynamic evolution of electrocatalytically induced processes with electrochemical liquid-phase electron microscopy (EM)is limited by the microcell configuration. Herein, a free-standing tri-layer graphene is integrated as a membrane and electrode material into the electrochemical chip and its suitability as a substrate electrode at the high cathodic potentials required for CO2 electroreduction (CO2ER) is evaluated. The three-layer stacked graphene is transferred onto an in-house fabricated single-working electrode chip for use with bulk-like reference and counter electrodes to facilitate evaluation of its effectiveness. Electrochemical measurements show that the graphene working electrode exhibits a wider inert cathodic potential range than the conventional glassy carbon electrode while achieving good charge transfer properties for nanocatalytic redox reactions. Operando scanning electron microscopy studies clearly demonstrate the improvement in spatial resolution but reveal a synergistic effect of the electron beam and the applied potential that limits the stability time window of the graphene-based electrochemical chip. By optimizing the operating conditions, in situ monitoring of Cu nanocube degradation is achieved at the CO2ER potential of -1.1V versus RHE. Thus, this improved microcell configuration allows EM observation of catalytic processes at potentials relevant to real systems.

Molecular dynamics simulation of the Brownian motion of biomolecules in liquid phase electron microscopy

Molecular dynamics simulation of the Brownian motion of biomolecules in liquid phase electron microscopy

Tri-layer graphene as membrane and electrode for liquid-phase electron microscopy studies of CO2 electroreduction nanocatalysts

Tri-layer graphene as membrane and electrode for liquid-phase electron microscopy studies of CO2 electroreduction nanocatalysts

Precision of Radiation Chemistry Networks: Playing Jenga with Kinetic Models for Liquid-Phase Electron Microscopy.

Liquid-phase transmission electron microscopy (LP-TEM) is a powerful tool to gain unique insights into dynamics at the nanoscale. The electron probe, however, can induce significant beam effects that often alter observed phenomena such as radiolysis of the aqueous phase. The magnitude of beam-induced radiolysis can be assessed by means of radiation chemistry simulations potentially enabling quantitative application of LP-TEM. Unfortunately, the computational cost of these simulations scales with the amount of reactants regarded. To minimize the computational cost, while maintaining accurate predictions, we optimize the parameter space for the solution chemistry of aqueous systems in general and for diluted HAuCl4 solutions in particular. Our results indicate that sparsened kinetic models can accurately describe steady-state formation during LP-TEM and provide a handy prerequisite for efficient multidimensional modeling. We emphasize that the demonstrated workflow can be easily generalized to any kinetic model involving multiple reaction pathways.

Crystal dissolution by particle detachment

Crystal dissolution, which is a fundamental process in both natural and technological settings, has been predominately viewed as a process of ion-by-ion detachment into a surrounding solvent. Here we report a mechanism of dissolution by particle detachment (DPD) that dominates in mesocrystals formed via crystallization by particle attachment (CPA). Using liquid phase electron microscopy to directly observe dissolution of hematite crystals — both compact rhombohedra and mesocrystals of coaligned nanoparticles — we find that the mesocrystals evolve into branched structures, which disintegrate as individual sub-particles detach. The resulting dissolution rates far exceed those for equivalent masses of compact single crystals. Applying a numerical generalization of the Gibbs-Thomson effect, we show that the physical drivers of DPD are curvature and strain inherently tied to the original CPA process. Based on the generality of the model, we anticipate that DPD is widespread for both natural minerals and synthetic crystals formed via CPA.

Applications ofelectron microscopic observations to electrochemistry inliquid electrolytes forbatteries.

Herein, we review notable points from observations of electrochemical reactions in a liquid electrolyte by liquid-phase electron microscopy. 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. Herein, we focus on two methods, of the liquid phase (scanning) transmission electron microscopy and liquid phase scanning electron microscopy, and summarize the advantages and disadvantages of both methods.

Tracking the Incorporation of Fe into NiO Electrocatalysts during Reaction with Liquid Phase Electron Microscopy and Time-Resolved Elemental Mapping.

Tracking the Incorporation of Fe into NiO Electrocatalysts during Reaction with Liquid Phase Electron Microscopy and Time-Resolved Elemental Mapping.

High-Throughput Low-Dose Biomolecule Imaging in Liquid Phase Electron Microscopy.

High-Throughput Low-Dose Biomolecule Imaging in Liquid Phase Electron Microscopy.

Tailoring the Acidity of Liquid Media with Ionizing Radiation: Rethinking the Acid–Base Correlation beyond pH

Advanced in situ techniques based on electrons and X-rays are increasingly used to gain insights into fundamental processes in liquids. However, probing liquid samples with ionizing radiation changes the solution chemistry under observation. In this work, we show that a radiation-induced decrease in pH does not necessarily correlate to an increase in acidity of aqueous solutions. Thus, pH does not capture the acidity under irradiation. Using kinetic modeling of radiation chemistry, we introduce alternative measures of acidity (radiolytic acidity π* and radiolytic ion product KW*), that account for radiation-induced alterations of both H+ and OH- concentration. Moreover, we demonstrate that adding pH-neutral solutes such as LiCl, LiBr, or LiNO3 can trigger a significant change in π*. This provides a huge parameter space to tailor the acidity for in situ experiments involving ionizing radiation, as present in synchrotron facilities or during liquid-phase electron microscopy.

An Iridium (III) complex revealing cytoskeleton nanostructures under super-resolution nanoscopy and liquid-phase electron microscopy

Live cell actin visualization is fundamental for exploring cellular motility, cytokinesis, intracellular transport, and other correlated functions. The current imaging techniques that allow imaging of actin in its native environment are optical and electron microscopy. Such imaging techniques offer high enough resolution to investigate the ultrastructure of actin however they come at the expense of actin integrity. Inspired by the lack of suitable probes that preserve actin's integrity, we designed a cyclometalated Ir (III) complex that interacts with live cells and displays light switch behaviour upon specific actin binding. The exceptional photophysical properties of the proposed probe allow unprecedented resolution of cytoskeleton ultrastructures under stimulated emission depletion (STED) super-resolution nanoscopy. Moreover, the Ir complex enables the capability of visualizing actin polymers and periodicity under correlative light electron microscopy (CLEM) and liquid-phase electron microscopy (LPEM) at ∼ 8 nm resolution.

Single Molecule Imaging with Liquid Phase Electron Microscopy

AbstractFew single‐molecule experiments have enabled the direct imaging of functional biomacromolecules in real‐time in their native liquid environments, resolving their conformational adaptations, transient interactions, and intermediate states. Liquid phase electron microscopy (LP‐EM), due to its unique combination of spatial and temporal resolution, has shown to be a promising tool. Recent experiments have enabled successful imaging of intact structures of organic molecules and biological systems with an ordinary electron microscope. Adapting image processing methods and quantitative data analysis from single particle experiments based on the optical microscope, quantifying motion and relaxation of these interacting molecules allows the experimental observations of pathways, to test theoretical predictions, and discovery of new mechanisms. Combining LP‐EM with tomography, fluorescence, and mass spectroscopy allows for probing multi‐dimensional structural and dynamic information. Challenges remain in obtaining high‐quality data in large quantities, which can be improved by developing new liquid cell platforms and machine learning‐based data analysis.

Non-classical crystallization of CeO2 by means of in situ electron microscopy.

During in situ liquid-phase electron microscopy (LP-EM) observations, the application of different irradiation dose rates may considerably alter the chemistry of the studied solution and influence processes, in particular growth pathways. While many processes have been studied using LP-EM in the last decade, the extent of the influence of the electron beam is not always understood and comparisons with corresponding bulk experiments are lacking. Here, we employ the radiolytic oxidation of Ce3+ in aqueous solution as a model reaction for the in situ LP-EM study of the formation of CeO2 particles. We compare our findings to the results from our previous study where a larger volume of Ce3+ precursor solution was subjected to γ-irradiation. We systematically analyze the effects of the applied irradiation dose rates and the induced diffusion of Ce ions on the growth mechanisms and the morphology of ceria particles. Our results show that an eight orders of magnitude higher dose rate applied during homogeneous electron-radiation in LP-EM compared to the dose rate using gamma-radiation does not affect the CeO2 particle growth pathway despite the significant higher Ce3+ to Ce4+ oxidation rate. Moreover, in both cases highly ordered structures (mesocrystals) are formed. This finding is explained by the stepwise formation of ceria particles via an intermediate phase, a signature of non-classical crystallization. Furthermore, when irradiation is applied locally using LP scanning transmission electron microscopy (LP-STEM), the higher conversion rate induces Ce-ion concentration gradients affecting the CeO2 growth. The appearance of branched morphologies is associated with the change to diffusion limited growth.

Deep learning-assisted analysis of single molecule dynamics from liquid-phase electron microscopy.

We apply U-Net and UNet++ to analyze single-molecule movies obtained from liquid-phase electron microscopy. Neural networks allow full automation, and high throughput analysis of these low signal-to-noise ratio images, while achieving higher segmentation accuracy, and avoiding subjective errors as compared to the conventional threshold methods. The analysis enables the quantification of transient dynamics in chemical systems and the capture of rare intermediate states by resolving local conformational changes within a single molecule.

Unraveling chemical processes during nanoparticle synthesis with liquid phase electron microscopy and correlative techniques.

Liquid phase transmission electron microscopy (LPTEM) has enabled unprecedented direct real time imaging of physicochemical processes during solution phase synthesis of metallic nanoparticles. LPTEM primarily provides images of nanometer scale, and sometimes atomic scale, metal nanoparticle crystallization processes, but provides little chemical information about organic surface ligands, metal-ligand complexes and reaction intermediates, and redox reactions. Likewise, complex electron beam-solvent interactions during LPTEM make it challenging to pinpoint the chemical processes, some involving exotic highly reactive radicals, impacting nanoparticle formation. Pairing LPTEM with correlative solution synthesis, ex situ chemical analysis, and theoretical modeling represents a powerful approach to gain a holistic understanding of the chemical processes involved in nanoparticle synthesis. In this feature article, we review recent work by our lab and others that has focused on elucidating chemical processes during nanoparticle synthesis using LPTEM and correlative chemical characterization and modeling, including mass and optical spectrometry, fluorescence microscopy, solution chemistry, and reaction kinetic modeling. In particular, we show how these approaches enable investigating redox chemistry during LPTEM, polymeric and organic capping ligands, metal deposition mechanisms on plasmonic nanoparticles, metal clusters and complexes, and multimetallic nanoparticle formation. Future avenues of research are discussed, including moving beyond electron beam induced nanoparticle formation by using light and thermal stimuli during LPTEM. We discuss prospects for real time LPTEM imaging and online chemical analysis of reaction intermediates using microfluidic flow reactors.