Taylor J. Woehl

Direct in Situ Determination of the Mechanisms Controlling Nanoparticle Nucleation and Growth

Although nanocrystal morphology is controllable using conventional colloidal synthesis, multiple characterization techniques are typically needed to determine key properties like the nucleation rate, induction time, growth rate, and the resulting morphology. Recently, researchers have demonstrated growth of nanocrystals by in situ electron beam reduction, offering direct observations of single nanocrystals and eliminating the need for multiple characterization techniques; however, they found nanocrystal morphologies consistent with two different growth mechanisms for the same electron beam parameters. Here we show that the electron beam current plays a role analogous to the concentration of reducing agent in conventional synthesis, by controlling the growth mechanism and final morphology of silver nanocrystals grown via in situ electron beam reduction. We demonstrate that low beam currents encourage reaction limited growth that yield nanocrystals with faceted structures, while higher beam currents encourage diffusion limited growth that yield spherical nanocrystals. By isolating these two growth regimes, we demonstrate a new level of control over nanocrystal morphology, regulated by the fundamental growth mechanism. We find that the induction threshold dose for nucleation is independent of the beam current, pixel dwell time, and magnification being used. Our results indicate that in situ electron microscopy data can be interpreted by classical models and that systematic dose experiments should be performed for all future in situ liquid studies to confirm the exact mechanisms underlying observations of nucleation and growth.

Experimental procedures to mitigate electron beam induced artifacts during in situ fluid imaging of nanomaterials

Scanning transmission electron microscopy of various fluid and hydrated nanomaterial samples has revealed multiple imaging artifacts and electron beam-fluid interactions. These phenomena include growth of crystals on the fluid stage windows, repulsion of particles from the irradiated area, bubble formation, and the loss of atomic information during prolonged imaging of individual nanoparticles. Here we provide a comprehensive review of these fluid stage artifacts, and we present new experimental evidence that sheds light on their origins in terms of experimental apparatus issues and indirect electron beam sample interactions with the fluid layer. A key finding is that many artifacts are a result of indirect electron beam interactions, such as production of reactive radicals in the water by radiolysis, and the associated crystal growth. The results presented here will provide a methodology for minimizing fluid stage imaging artifacts and acquiring quantitative in situ observations of nanomaterial behavior in a liquid environment.

Direct in Situ Observation of Nanoparticle Synthesis in a Liquid Crystal Surfactant Template

Controlled and reproducible synthesis of tailored materials is essential in many fields of nanoscience. In order to control synthesis, there must be a fundamental understanding of nanostructure evolution on the length scale of its features. Growth mechanisms are usually inferred from methods such as (scanning) transmission electron microscopy ((S)TEM), where nanostructures are characterized after growth is complete. Such post mortem analysis techniques cannot provide the information essential to optimize the synthesis process, because they cannot measure nanostructure development as it proceeds in real time. This is especially true in the complex rheological fluids used in preparation of nanoporous materials. Here we show direct in situ observations of synthesis in a highly viscous lyotropic liquid crystal template on the nanoscale using a fluid stage in the STEM. The nanoparticles nucleate and grow to ∼5 nm particles, at which point growth continues through the formation of connections with other nanoparticles around the micelles to form clusters. Upon reaching a critical size (>10-15 nm), the clusters become highly mobile in the template, displacing and trapping micelles within the growing structure to form spherical, porous nanoparticles. The final products match those synthesized in the lab ex situ. This ability to directly observe synthesis on the nanoscale in rheological fluids, such as concentrated aqueous surfactants, provides an unprecedented understanding of the fundamental steps of nanomaterial synthesis. This in turn allows for the synthesis of next-generation materials that can strongly impact important technologies such as organic photovoltaics, energy storage devices, catalysis, and biomedical devices.

Factors influencing quantitative liquid (scanning) transmission electron microscopy

One of the experimental challenges in the study of nanomaterials in liquids in the (scanning) transmission electron microscope ((S)TEM) is gaining quantitative information. A successful experiment in the fluid stage will depend upon the ability to plan for sensitive factors such as the electron dose applied, imaging mode, acceleration voltage, beam-induced solution chemistry changes, and the specifics of solution reactivity. In this paper, we make use of a visual approach to show the extent of damage of different instrumental and experimental factors in liquid samples imaged in the (S)TEM. Previous results as well as new insights are presented to create an overview of beam-sample interactions identified for changing imaging and experimental conditions. This work establishes procedures to understand the effect of the electron beam on a solution, provides information to allow for a deliberate choice of the optimal experimental conditions to enable quantification, and identifies the experimental factors that require further analysis for achieving fully quantitative results in the liquid (S)TEM.

Nucleation of Iron Oxide Nanoparticles Mediated by Mms6 Protein in Situ

Biomineralization proteins are widely used as templating agents in biomimetic synthesis of a variety of organic-inorganic nanostructures. However, the role of the protein in controlling the nucleation and growth of biomimetic particles is not well understood, because the mechanism of the bioinspired reaction is often deduced from ex situ analysis of the resultant nanoscale mineral phase. Here we report the direct visualization of biomimetic iron oxide nanoparticle nucleation mediated by an acidic bacterial recombinant protein, Mms6, during an in situ reaction induced by the controlled addition of sodium hydroxide to solution-phase Mms6 protein micelles incubated with ferric chloride. Using in situ liquid cell scanning transmission electron microscopy we observe the liquid iron prenucleation phase and nascent amorphous nanoparticles forming preferentially on the surface of protein micelles. Our results provide insight into the early steps of protein-mediated biomimetic nucleation of iron oxide and point to the importance of an extended protein surface during nanoparticle formation.

Minimum Cost Multi-Way Data Association for Optimizing Multitarget Tracking of Interacting Objects

This paper presents a general formulation for a minimum cost data association problem which associates data features via one-to-one, m-to-one and one-to-n links with minimum total cost of the links. A motivating example is a problem of tracking multiple interacting nanoparticles imaged on video frames, where particles can aggregate into one particle or a particle can be split into multiple particles. Many existing multitarget tracking methods are capable of tracking non-interacting targets or tracking interacting targets of restricted degrees of interactions. The proposed formulation solves a multitarget tracking problem for general degrees of inter-object interactions. The formulation is in the form of a binary integer programming problem. We propose a polynomial time solution approach that can obtain a good relaxation solution of the binary integer programming, so the approach can be applied for multitarget tracking problems of a moderate size (for hundreds of targets over tens of time frames). The resulting solution is always integral and obtains a better duality gap than the simple linear relaxation solution of the corresponding problem. The proposed method was validated through applications to simulated multitarget tracking problems and a real multitarget tracking problem.

Correlative electron and fluorescence microscopy of magnetotactic bacteria in liquid: toward in vivo imaging.

Magnetotactic bacteria biomineralize ordered chains of uniform, membrane-bound magnetite or greigite nanocrystals that exhibit nearly perfect crystal structures and species-specific morphologies. Transmission electron microscopy (TEM) is a critical technique for providing information regarding the organization of cellular and magnetite structures in these microorganisms. However, conventional TEM can only be used to image air-dried or vitrified bacteria removed from their natural environment. Here we present a correlative scanning TEM (STEM) and fluorescence microscopy technique for imaging viable cells of Magnetospirillum magneticum strain AMB-1 in liquid using an in situ fluid cell TEM holder. Fluorescently labeled cells were immobilized on microchip window surfaces and visualized in a fluid cell with STEM, followed by correlative fluorescence imaging to verify their membrane integrity. Notably, the post-STEM fluorescence imaging indicated that the bacterial cell wall membrane did not sustain radiation damage during STEM imaging at low electron dose conditions. We investigated the effects of radiation damage and sample preparation on the bacteria viability and found that approximately 50% of the bacterial membranes remained intact after an hour in the fluid cell, decreasing to ~30% after two hours. These results represent a first step toward in vivo studies of magnetite biomineralization in magnetotactic bacteria.

Understanding the Role of Solvation Forces on the Preferential Attachment of Nanoparticles in Liquid

Optimization of colloidal nanoparticle synthesis techniques requires an understanding of underlying particle growth mechanisms. Nonclassical growth mechanisms are particularly important as they affect nanoparticle size and shape distributions, which in turn influence functional properties. For example, preferential attachment of nanoparticles is known to lead to the formation of mesocrystals, although the formation mechanism is currently not well-understood. Here we employ in situ liquid cell scanning transmission electron microscopy and steered molecular dynamics (SMD) simulations to demonstrate that the experimentally observed preference for end-to-end attachment of silver nanorods is a result of weaker solvation forces occurring at rod ends. SMD reveals that when the side of a nanorod approaches another rod, perturbation in the surface-bound water at the nanorod surface creates significant energy barriers to attachment. Additionally, rod morphology (i.e., facet shape) effects can explain the majority of the side attachment effects that are observed experimentally.

Peptide-Directed PdAu Nanoscale Surface Segregation: Toward Controlled Bimetallic Architecture for Catalytic Materials

Bimetallic nanoparticles are of immense scientific and technological interest given the synergistic properties observed when two different metallic species are mixed at the nanoscale. This is particularly prevalent in catalysis, where bimetallic nanoparticles often exhibit improved catalytic activity and durability over their monometallic counterparts. Yet despite intense research efforts, little is understood regarding how to optimize bimetallic surface composition and structure synthetically using rational design principles. Recently, it has been demonstrated that peptide-enabled routes for nanoparticle synthesis result in materials with sequence-dependent catalytic properties, providing an opportunity for rational design through sequence manipulation. In this study, bimetallic PdAu nanoparticles are synthesized with a small set of peptides containing known Pd and Au binding motifs. The resulting nanoparticles were extensively characterized using high-resolution scanning transmission electron microscopy, X-ray absorption spectroscopy, and high-energy X-ray diffraction coupled to atomic pair distribution function analysis. Structural information obtained from synchrotron radiation methods was then used to generate model nanoparticle configurations using reverse Monte Carlo simulations, which illustrate sequence dependence in both surface structure and surface composition. Replica exchange with solute tempering molecular dynamics simulations were also used to predict the modes of peptide binding on monometallic surfaces, indicating that different sequences bind to the metal interfaces via different mechanisms. As a testbed reaction, electrocatalytic methanol oxidation experiments were performed, wherein differences in catalytic activity are clearly observed in materials with identical bimetallic composition. Taken together, this study indicates that peptides could be used to arrive at bimetallic surfaces with enhanced catalytic properties, which could be leveraged for rational bimetallic nanoparticle design using peptide-enabled approaches.

Visualization of iron-binding micelles in acidic recombinant biomineralization protein, MamC

Biological macromolecules are utilized in low-temperature synthetic methods to exert precise control over nanoparticle nucleation and placement. They enable low-temperature formation of a variety of functional nanostructured materials with properties often not achieved via conventional synthetic techniques. Here we report on the in situ visualization of a novel acidic bacterial recombinant protein, MamC, commonly present in the magnetosome membrane of several magnetotactic bacteria, including Magnetococcus marinus, strain MC-1. Our findings provide an insight into the self-assembly of MamC and point to formation of the extended protein surface, which is assumed to play an important role in the formation of biotemplated inorganic nanoparticles. The self-organization of MamC is compared to the behavior of another acidic recombinant iron-binding protein, Mms6.