Models of cation exchange mechanisms and driving forces have proven effective predictors of clay behavior and chemistry, but are largely theoretical, particularly in complex systems involving high ionic strength brines or systems where hydration is controlled by relative humidity. In arid and cold environments, such as Mars, cyclical relative humidity variations may play a role in chemical alteration, particularly if clay minerals such as smectite are in the presence of salts. This study examines the effects of relative humidity on smectite-salt mixtures using environmental scanning electron microscopy (ESEM) to observe the physiochemical effects of salt deliquescence and desiccation on smectite textures and elemental distributions. Results demonstrate that even reaction periods as short as a few minutes allow ample time for relative humidity to affect the smectite-salt mixtures. In addition to smectite swelling and salt deliquescence, we also observed rapid changes in element distributions within the smectite and new crystal growth in the presence of high relative humidity. Even in the absence of bulk liquid water, exchangeable cations migrated out of the smectite and formed new crystals at the smectite-salt interface. The observed microscopic changes in elemental distributions indicate that the migration of cations driven by cation exchange led to secondary mineral precipitation, likely a CaSO4 mineral, within a sub-micrometer-thick layer of water on the smectite grains. The results of this study demonstrate that during periods of elevated relative humidity, active smectite mineral alteration and secondary mineral precipitation may be possible on present-day Mars where salts and smectites are in direct physical contact.

An experimental study of the swelling anisotropy of the Callovo-Oxfordian argillaceous rock under hydration is presented. The investigation, which combines environmental scanning electron microscopy (ESEM) and digital image correlation techniques, has been carried out at the micrometric scale of the composite microstructure of the rock. Specimens were hydrated in the ESEM over a wide range of relative humidity and observations conducted on two planes: plane 1 parallel to the bedding plane, and plane 2 perpendicular to it. The observations reveal that the local swelling (which can be quantified at a local gauge length of about 5 μm) is strongly anisotropic in both planes. The global swelling, measured over areas of about 500 μm in width, is also clearly anisotropic in plane 2 (with major swelling direction normal to the bedding plane), but not in plane 1. The global isotropy in plane 1 arises from the uniform distribution of the orientation of anisotropic local strains, while the anisotropic swelling in plane 2 is due to a preferred local orientation.

Drug release from oral pharmaceutical formulations can be modified by applying a polymeric coating film with controlled mass transport properties. Interaction of the coating film with water may crucially influence its composition and permeability to both water and drug. Understanding this interaction between film microstructure, wetting, and mass transport is important for the development of new coatings. We present a novel method for controlled wetting of polymer coating films in an environmental scanning electron microscope, providing direct visual information about the processes occurring as the film goes from dry to wet. Free films made of phase-separated blends of water-insoluble ethyl cellulose (EC) and water-soluble hydroxypropyl cellulose (HPC) were used as a model system, and the blend ratio was varied to study the effect on the water transport properties. Local variations in water transport through the EC/HPC films were directly observed, enabling the immediate analysis of the structure–mass transport relationships. The leaching of HPC could be studied by evaporating water from the films in situ. Significant differences were observed between films of varying composition. The method provides a valuable complement to the current approach of making distinct diffusion and microscopy experiments for studying the dynamic interaction of polymer films with water.

Scanning transmission electron microscopy (STEM) of specimens in liquid, so-called Liquid STEM, is capable of imaging the individual subunits of macromolecular complexes in whole eukaryotic cells in liquid. This paper discusses this new microscopy modality within the context of state-of-the-art microscopy of cells. The principle of operation and equations for the resolution are described. The obtained images are different from those acquired with standard transmission electron microscopy showing the cellular ultrastructure. Instead, contrast is obtained on specific labels. Images can be recorded in two ways, either via STEM at 200 keV electron beam energy using a microfluidic chamber enclosing the cells, or via environmental scanning electron microscopy at 30 keV of cells in a wet environment. The first series of experiments involved the epidermal growth factor receptor labeled with gold nanoparticles. The labels were imaged in whole fixed cells with nanometer resolution. Since the cells can be kept alive in the microfluidic chamber, it is also feasible to detect the labels in unfixed, live cells. The rapid sample preparation and imaging allows studies of multiple whole cells.

The size of gold nanoparticles (AuNPs) can influence various aspects of their cellular uptake. Light microscopy is not capable of resolving most AuNPs, while electron microscopy (EM) is not practically capable of acquiring the necessary statistical data from many cells and the results may suffer from various artifacts. Here, we demonstrate the use of a fast EM method for obtaining high-resolution data from a much larger population of cells than is usually feasible with conventional EM. A549 (human lung carcinoma) cells were subjected to uptake protocols with 10, 15, or 30 nm diameter AuNPs with adsorbed serum proteins. After 20 min, 24 h, or 45 h, the cells were fixed and imaged in whole in a thin layer of liquid water with environmental scanning electron microscopy equipped with a scanning transmission electron microscopy detector. The fast preparation and imaging of 145 whole cells in liquid allowed collection of nanoscale data within an exceptionally small amount of time of ~80 h. Analysis of 1,041 AuNP-filled vesicles showed that the long-term AuNP storing lysosomes increased their average size by 80 nm when AuNPs with 30 nm diameter were uptaken, compared to lysosomes of cells incubated with AuNPs of 10 and 15 nm diameter.

Environmental scanning electron microscopy has been extensively used for studying the wetting properties of different materials. For some types of investigation, however, the traditional ways of conducting in situ dynamic wetting experiments do not offer sufficient control over the wetting process. Here, we present a novel method for controlled wetting of materials in the environmental scanning electron microscope (ESEM). It offers improved control of the point of interaction between the water and the specimen and renders it more accessible for imaging. It also enables the study of water transport through a material by direct imaging. The method is based on the use of a piezo-driven nanomanipulator to bring a specimen in contact with a water reservoir in the ESEM chamber. The water reservoir is established by local condensation on a Peltier-cooled surface. A fixture was designed to make the experimental setup compatible with the standard Peltier cooling stage of the microscope. The developed technique was successfully applied to individual cellulose fibers, and the absorption and transport of water by individual cellulose fibers were imaged.

The degradation of Sydney sandstone used to build the heritage St Mary's Cathedral in Sydney, Australia, has been investigated using environmental scanning electron microscopy combined with energy dispersive X-ray spectroscopy. This technique provided the structural details of the cementing clay and an elemental characterization the sandstone. The observed differences in the elemental composition of the unweathered and weathered sandstones were associated with changes to the clay microstructure upon weathering. The results support the substitution theory that Fe3+ replaces Al3+ in the kaolinite clay component upon weathering. An examination of the impurities present prior to a nonstructural iron removal treatment revealed the presence of minerals that may provide a source of the elements responsible for the substitution process.

Testate amoebae (TA) are a group of free-living protozoa, important in ecology and paleoecology. Testate amoebae taxonomy is mainly based on the morphological features of the shell, as examined by means of light microscopy or (environmental) scanning electron microscopy (SEM/ESEM). We explored the potential applications of confocal laser scanning microscopy (CLSM), two photon excitation microscopy (TPEM), phase contrast, differential interference contrast (DIC Nomarski), and polarization microscopy to visualize TA shells and inner structures of living cells, which is not possible by SEM or environmental SEM. Images captured by CLSM and TPEM were utilized to create three-dimensional (3D) visualizations and to evaluate biovolume inside the shell by stereological methods, to assess the function of TA in ecosystems. This approach broadens the understanding of TA cell and shell morphology, and inner structures including organelles and endosymbionts, with potential implications in taxonomy and ecophysiology.

Quantitative chemical analysis by energy-dispersive X-ray spectrometry (EDS) in the environmental scanning electron microscope (ESEM) is difficult. This analysis is complicated by the spread of the electron beam by chamber gas molecules and the necessity for surface charge neutralization. Without charge neutralization, errors in quantitative analysis can range up to 15–20% relative. It is possible to achieve the error expected of traditional EDS, ±5% relative error, using a newly developed surface charge neutralization scheme for the ESEM. Estimates of accuracy and precision are based on studies of the National Bureau of Standards (now National Institutes for Science and Technology) Standard Reference Material 482, a series of certified copper–gold alloys. The scheme for charge neutralization requires an independent path to ground at or near the surface of the specimen. The current through the ground path must be maintained at zero by adjusting the voltage on the Gaseous Secondary Electron DetectorTM when the sample chamber is at a gas pressure of 1–2 torr. This procedure forms the exact number of chamber gas positive ions to neutralize negative electrical charge on the specimen surface from electron bombardment.

Energy dispersive X-ray spectrometry of uncoated insulators performed at low beam energy (incident energy ≤ 5 keV) and in the variable pressure scanning electron microscope and the environmental scanning electron microscope is subject to spectral artifacts. Charging decelerates the incident beam electrons and reduces the impact energy, lowering the available overvoltage to excite characteristic X-ray peaks. The Duane–Hunt limit of the X-ray bremsstrahlung continuum is commonly used as a diagnostic of charging. Dynamic charging effects can hide the true impact of charging on the X-ray spectrum. Careful examination of the behavior of the X-ray spectrum with time and other variables is needed to avoid spectral artifacts, particularly on relative X-ray intensities.

In variable pressure scanning electron microscopy (VPSEM) the current data suggests that considerable caution is required in the interpretation of X-ray data from nonconductive samples, depending on the operating conditions. This article reviews some of the documented approaches and presents data that illustrate the nature and magnitude of the effects of charge above, on, and in the sample on the detected X-ray emissions from the sample and from elsewhere within the VPSEM specimen chamber. The collection of reliable and reproducible X-ray data has been found to require relatively high specimen chamber gas pressures, at the upper end of or beyond the available pressures for most VPSEMs. It is also shown that sample characteristics, including composition, strongly influence local charge effects, which can significantly affect the primary electron landing energy and consequently the resultant emitted X-ray signal under low pressure environments.

The Bugscope project is an educational outreach program for kindergarten to grade 12 (K–12) classrooms. The project provides a resource to classrooms so that they may remotely operate a scanning electron microscope to image insects at high magnification. The microscope is remotely controlled in real time from a classroom computer over the Internet using a Web browser. Bugscope provides a state-of-the-art microscope resource for teachers that can be readily integrated into classroom activities. The Bugscope project provides a low-cost, sustainable model for research groups to support K–12 education outreach projects.

In large bandgap semiconductors and insulators, the threshold energies for e–h pair production and ionization damage can lie above the vacuum level. For low energy imaging, a window is then opened whose width is potentially sensitive to local changes in work function, doping level, or acidity. Recent progress and future opportunities for damage-free imaging of these properties using low energy electrons are discussed in the light of the underlying physics, as well as of recent instrumental developments in low energy electron microscopy (LEEM), environmental scanning electron microscopy (ESEM), photoelectron emission microscopy (PEEM), scanned probe microscopy (SPM), and projection electron microscopy.