The Nanoaquarium: A Nanofluidic Platform for in SiTu Transmission Electron Microscopy in Liquid Media

There are many scientifically interesting and technologically relevant nanoscale phenomena that take place in liquid media. Examples include aggregation and assembly of nanoparticles; colloidal crystal formation; liquid phase growth of structures such as nanowires; electrochemical deposition and etching for fabrication processes and battery applications; interfacial phenomena; boiling and cavitation; and biological interactions. Understanding of these fields would benefit greatly from real-time, in situ transmission electron microscope (TEM) imaging with nanoscale resolution. Most liquids cannot be imaged by traditional TEM due to evaporation in the high vacuum environment and the requirement that samples be very thin. Liquid-cell in situ TEM has emerged as an exciting new experimental technique that hermetically seals a thin slice of liquid between two electron transparent membranes to enable TEM imaging of liquid-based processes. This work presents details of the fabrication of a custom-made liquid-cell in situ TEM device, dubbed the nanoaquarium. The nanoaquarium’s highlights include an exceptionally thin sample cross section (10s to 100s of nm); wafer scale processing that enables high-yield mass production; robust hermetic sealing that provides leak-free operation without use of glue, epoxy, or any polymers; compatibility with lab-on-chip technology; and on-chip integrated electrodes for sensing and actuation. The fabrication process is described, with an emphasis on direct wafer bonding. Experimental results involving direct observation of colloid aggregation using an aqueous solution of gold nanoparticles are presented. Quantitative analysis of the growth process agrees with prior results and theory, indicating that the experimental technique does not radically alter the observed phenomenon. For the first time, in situ observations of nanoparticles at a contact line and in an evaporating thin film of liquid are reported, with applications for techniques such as dip-coating and drop-casting, commonly used for depositing nanoparticles on a surface via convective-capillary assembly. Theoretical analysis suggests that the observed particle motion and aggregation are caused by gradients in surface tension and disjoining pressure in the thin liquid film

A Nanoaquarium for in situ Electron Microscopy in Liquid Media

The understanding of many nanoscale processes occurring in liquids such as colloidal crystal formation, aggregation, nanowire growth, electrochemical deposition, and biological interactions would benefit greatly from real-time, in situ imaging with the nanoscale resolution of transmission electron microscopes (TEMs) and scanning transmission electron microscopes (STEMs). However, these imaging tools cannot readily be used to observe processes occurring in liquid media without addressing two experimental hurdles: sample thickness and sample evaporation in the high vacuum microscope chamber. To address these challenges, we have developed a nano-Hele-Shaw cell, dubbed the nanoaquarium. The device consists of a hermetically-sealed, 100 nm tall, liquid-filled chamber sandwiched between two freestanding, 50 nm thick, silicon nitride membranes. Embedded electrodes are integrated into the device. This fluid dynamics video features particle motion and aggregation during in situ STEM of nanoparticles suspended in liquids. The first solution contains 5 nm gold particles, 50 nm gold particles and 50 nm polystyrene particles in water. The second solution contains 5 nm gold particles in water. The imaging was carried out with a FEI Quanta 600 FEG Mark II with a STEM detector. In the footage of the multi-particle solution, note that the 50 nm gold particles prominently decorate the clusters and are clearly distinguished. In the footage of the 5 nm gold particles, diffusion-limited aggregation is observed. Individual particles and small clusters are seen diffusing throughout the field of view, bumping into each other and bonding irreversibly to form a fractal structure. The rate of aggregation and the fractal dimension of the aggregates are consistent with light scattering measurements, indicating that the electron beam does not greatly alter the observed phenomenon.Comment: videos are include

\u3cem\u3eIn Situ\u3c/em\u3e Liquid-Cell Electron Microscopy of Colloid Aggregation and Growth Dynamics

We report on real-time observations of the aggregation of gold nanoparticles using a custom-made liquid cell that allows for in situ electron microscopy. Process kinetics and fractal dimension of the aggregates are consistent with three-dimensional cluster-cluster diffusion-limited aggregation, even for large aggregates, for which confinement effects are expected. This apparent paradox was resolved through in situ observations of the interactions between individual particles as well as clusters at various stages of the aggregation process that yielded the large aggregates. The liquid cell described herein facilitates real-time observations of various processes in liquid media with the high resolution of the electron microscope

Bubble and Pattern Formation in Liquid Induced by an Electron Beam

Liquid cell electron microscopy has emerged as a powerful technique for in situ studies of nanoscale processes in liquids. An accurate understanding of the interactions between the electron beam and the liquid medium is essential to account for, suppress, and exploit beam effects. We quantify the interactions of high energy electrons with water, finding that radiolysis plays an important role, while heating is typically insignificant. For typical imaging conditions, we find that radiolysis products such as hydrogen and hydrated electrons achieve equilibrium concentrations within seconds. At sufficiently high dose-rate, the gaseous products form bubbles. We image bubble nucleation, growth, and migration. We develop a simplified reaction-diffusion model for the temporally and spatially varying concentrations of radiolysis species and predict the conditions for bubble formation by . We discuss the conditions under which hydrated electrons cause precipitation of cations from solution, and show that the electron beam can be used to “write” structures directly, such as nanowires and other complex patterns, without the need for a mask

Women Scholars, Integration, and the Marianist Tradition: Learning From Our Culture and Ourselves

In the fall of 1997, a group of junior tenure-track women faculty in the Department of Teacher Education at the University of Dayton decided to meet regularly in order to support each others’ scholarly endeavors in the process of achieving promotion and tenure. The group of subsequently became known as the Writing ”“Writers’ Support Group (WWSG). In 2000, the group conducted a self-study of its group process to determine how the formation of women’s WWSG fit with the mission and characteristics of a Marianist university. The results suggest that, although each of the characteristics could be identified in the group processes, the group best identified with the Marianist mandate to educate in family spirit. Each member of the group considered the possible reasons for this outcome

Women Scholars, Integration, and the Marianist Tradition: Learning from our Culture and Ourselves

In the fall of 1997, a group of junior tenure-track women faculty in the Department of Teacher Education at the University of Dayton decided to meet regularly in order to support each other’s scholarly endeavors in the process of achieving promotion and tenure. The group of subsequently became known as the Writing-Writers’ Support Group (WWSG). In 2000, the group conducted a self-study of its group process to determine how the formation of women’s WWSG fit with the mission and characteristics of a Marianist university. The results suggest that, although each of the characteristics could be identified in the group processes, the group best identified with the Marianist mandate to educate in family spirit. Each member of the group considered the possible reasons for this outcome

The Effect of Epstein-Barr Virus Latent Membrane Protein 2 Expression on the Kinetics of Early B Cell Infection

Infection of human B cells with wild-type Epstein-Barr virus (EBV) in vitro leads to activation and proliferation that result in efficient production of lymphoblastoid cell lines (LCLs). Latent Membrane Protein 2 (LMP2) is expressed early after infection and previous research has suggested a possible role in this process. Therefore, we generated recombinant EBV with knockouts of either or both protein isoforms, LMP2A and LMP2B (Δ2A, Δ2B, Δ2A/Δ2B) to study the effect of LMP2 in early B cell infection. Infection of B cells with Δ2A and Δ2A/Δ2B viruses led to a marked decrease in activation and proliferation relative to wild-type (wt) viruses, and resulted in higher percentages of apoptotic B cells. Δ2B virus infection showed activation levels comparable to wt, but fewer numbers of proliferating B cells. Early B cell infection with wt, Δ2A and Δ2B viruses did not result in changes in latent gene expression, with the exception of elevated LMP2B transcript in Δ2A virus infection. Infection with Δ2A and Δ2B viruses did not affect viral latency, determined by changes in LMP1/Zebra expression following BCR stimulation. However, BCR stimulation of Δ2A/Δ2B cells resulted in decreased LMP1 expression, which suggests loss of stability in viral latency. Long-term outgrowth assays revealed that LMP2A, but not LMP2B, is critical for efficient long-term growth of B cells in vitro. The lowest levels of activation, proliferation, and LCL formation were observed when both isoforms were deleted. These results suggest that LMP2A appears to be critical for efficient activation, proliferation and survival of EBV-infected B cells at early times after infection, which impacts the efficient long-term growth of B cells in culture. In contrast, LMP2B did not appear to play a significant role in these processes, and long-term growth of infected B cells was not affected by the absence of this protein. © 2013 Wasil et al