The Microscopy and Imaging Center at Texas A&M University is a multi-user facility involved with preparation and analysis of many different biological and materials sciences projects. Vapor stabilization and coating is an important part of our specimen preparation methodology for difficult biological and materials, especially polymer, samples. The procedure for all our vapor preparation techniques is done in a simple, economical apparatus set up in a properly functioning fume hood with a flow rate of at least 100 ft/min (Fig. 1). The apparatus is made from a glass petri dish or a glass petri dish for the bottom and an appropriate size beaker for the top. Specimens, mounted on stubs, are placed inside the chamber and the fixative (osmium tetroxide, ruthenium tetroxide or acrolein) is placed in a small container (plastic bottle cap) near the specimens.

X-ray ultramicroscopy in the SEM is a relatively new application in the wider field of X-ray microscopy. This latter field includes synchrotron and cabinet-based systems that vary in their X-ray power, capability, sample size, spatial resolution, and convenience. One important capability of the SEM-hosted X-ray microscope is that the normal SEM imaging and analytical functions such as secondary and backscattered imaging and microanalysis by EDX or WDS are unimpeded. X-ray imaging then serves as a complement to the normal use of the SEM. The convenience of easy access in an SEM lab to an X-ray microscope with 3D tomographic capability makes this an important development.

Hyperspectral imaging (also known as spectrum imaging) requires software for extracting the signatures present in every spectrum. However, commercial software available for spectrum analysis remains expensive, complicated, and often not transparent regarding the internal workings and approximations made. For user facilities, educational institutes, and other settings where multiple users on a single tool can be expected, the limited availability of software becomes the bottleneck to data analysis, user training, and throughput. The Cornell Spectrum Imager (CSI) was developed as a universal data analysis tool to be freely distributed, to run on all computers, and to minimize training. This is accomplished by using one simple interface for imaging, cathodoluminescence, Raman, Electron Energy Loss Spectroscopic (EELS), and EDX data analysis. This article demonstrates the CSI plugins for ImageJ by guiding you through the basic workflow for processing EELS maps.

The continued focus on improving materials, combined with the fact that it is now commonly understood that material properties are affected by characteristics at the atomic level, give rise to the need to characterize and image at the best resolutions possible. The (Scanning) Transmission Electron Microscope ((S)TEM) has the capability to image structures with atomic resolution and provides, at the same time, information on the chemical composition, bonding and electronic structure of the material. The nanoresearcher's continued need for the ultimate resolution has accelerated the development of next generation electron optics and technology.

One of the main obstacles to performing electron crystallography analysis in a TEM is that the acquired electron diffraction data often exhibits some form of distortion introduced by the lens system. Recognizing this problem, Capitani et al. has proposed a method to detect such distortion, which is primarily elliptical, by using a single crystal standard. Once such elliptical distortion is characterized, electron diffraction data acquired later can then be corrected by means of image processing. However, it may be desirable to correct such distortion at the instrument level. In this article, a different approach to measuring diffraction elliptical distortion is proposed by characterizing diffraction ring patterns and it is demonstrated that by varying the objective lens stigmation settings, it is possible to eliminate this elliptical distortion completely.

Electrochemical and electrocatalytic phenomena underpin a broad spectrum of energy, chemical, and information technologies. Examples range from electrocatalytic activation of oxygen reduction reactions in fuel cells, gas-solid reactions, and ionic transport in gas sensors and oxygen pumps, to a wide gamut of electrochemical and transport phenomena in primary and secondary batteries. Equally important is the role of ionic phenomena in information technologies, as exemplified by a recent wave of interest in memristive and electroresisitive information storage and logic devices. This article examines a new type of scanning probe microscopy (SPM), referred to as electrochemical strain microscopy (ESM), that can provide a better understanding of these complex phenomena.

Since the early 1990s EBSD (Electron Back Scatter Diffraction) has been developed to become a mature crystallographic analysis technique at the micro and nano-scale. It is applied in a SEM (Scanning Electron Microscope) on samples with a very smooth and clean surface. It provides quantitative orientations of individual grains, and by mapping a larger area, multiple grain analysis, texture, and grain boundaries can be examined. As the useful information for the EBSD technique comes from a very shallow depth in the material (10 to 20nm), it is a surface analysis technique showing lateral 2D distributions of crystal orientations.

With advancements in aberration-corrected electron optics, the resolution in scanning transmission electron microscopy (STEM) has been significantly improved. More importantly, the reduction of the probe size and the increase of the probe current density enable the acquisition of elemental maps at the atomic scale in a fast manner using both EELS and EDS. With the latest generations of EELS spectrometers, atomic resolution compositional and chemical maps allow interfaces, oxidation state, and even single atoms to be examined with increasing detail. These improvements have also enabled elemental and chemical maps to be acquired rapidly using both low- and high-energy edges from elements across the periodic table, including heavy atoms such Au or Pt. On the EDS side, the introduction of large-area silicon drift detectors (SDD) has allowed these high-beam-current sources to be fully utilized. Improved detector area and support for higher count rates, compared to the previous generation of EDS detectors, allows the acquisition of EDS intensity maps from most types of material in the STEM, even for moderately thin samples. For the case of light elements, the reduced detector dead layer of SDD-based systems partially overcomes the very low florescence yield typical of low-energy X-ray lines. Also as reported in, atomic-level X-ray maps using fast detectors and bright sources can now be collected under some conditions. Because EDS and EELS provide complementary information about the sample, and are both generated with the electron beam, it would be wasteful to not acquire both datasets with every sample run.

A microscope, by definition, provides structural and analytical information about objects that are too small to see with the unaided eye. From the very first microscope, efforts to improve its capabilities and push them to ever-finer length scales have been pursued. In this context, it would seem that the concept of an ultimate microscope would have received much attention by now; but has it really ever been defined? Human knowledge extends to structures on a scale much finer than atoms, so it might seem that a proton-scale microscope or a quark-scale microscope would be the ultimate. However, we argue that an atomic-scale microscope is the ultimate for the following reason: the smallest building block for either synthetic structures or natural structures is the atom. Indeed, humans and nature both engineer structures with atoms, not quarks. So far as we know, all building blocks (atoms) of a given type are identical; it is the assembly of the building blocks that makes a useful structure (see Figure 1). Thus, would a microscope that determines the position and identity of every atom in a structure with high precision and for large volumes be the ultimate microscope? We argue, yes. In this article, we consider how it could be built, and we ponder the answer to the equally important follow-on questions: who would care if it is built, and what could be achieved with it?

Coupling between electrical and mechanical phenomena is an important feature of functional inorganic materials and biological systems alike. The applications of electromechanically active materials include sonar, ultrasonic and medical imaging, sensors, actuators, and energy-harvesting technologies, as well as non-volatile computer memories. Electromechanical coupling in electromotor proteins and cellular membranes is the universal basis for biological functionalities from hearing to cardiac activity. The future will undoubtedly see the emergence of broad arrays of piezoelectric, biological, and molecular-based electromechanical systems to allow mankind the capability not only to “think” but also “act” on the nanoscale. The need for probing electromechanical functionalities has led to the development of Piezoresponse Force Microscopy (PFM) as a tool for local nanoscale imaging (Figures 1 and 2), spectroscopy, and manipulation of piezoelectric and ferroelectric materials.

2D-crystallization in combination with transmission electron microscopy (TEM) is one of the few methods for the structural analysis of membrane proteins in their native state. However, the parameters for the generation of large crystalline sheets are typically difficult to identify for a given protein. Many repetitive and time consuming screening steps by TEM are therefore necessary to find the best crystallization and preparation conditions. Although several software packages offer the possibility to control an electron microscope, none is completely adapted for a fully automated and completely integrated acquisition and analysis of 2D crystals. Here we report on the development of a fully automatic screening and on-line analysis software for the fast and automatic survey of large quantities of negatively stained EM samples for 2D crystallography.

Capabilities for in-situ studies of materials at elevated temperatures and under gaseous environments have received increasing attention in recent years [1]. With the advent of electron microscopes that provide routine imaging at the atomic level (e.g. aberration-corrected TEM and STEM instruments), it is of particular interest to be able to record images at high temperatures while retaining the inherent resolution of the microscope; that is, the resolution is not limited by drift in the heating holder or other instabilities associated with its operation. A number of commercial and experimental heating devices have been used over the years; some holders are designed with miniature furnaces that heat entire grids [2], while a more recent development used a tiny spiral filament coated with a carbon film as the heater element [3]. These devices, while very useful for some applications (particularly in “environmental microscopes” that employ differential pumping to allow gases at some elevated pressure to be injected around the specimen), are invariably not as stable as might be desired for sub-Ångström imaging experiments. They are also limited by the speed at which the sample can be heated to temperature for stable operation. In collaboration with Protochips Inc. (Raleigh, NC), our laboratory is developing a novel new technology for in-situ heating experiments that overcomes a number of performance problems associated with standard heating stage technologies [4].

Our group studies the spread of cells in primary breast tumors to distant sites, a process that is called metastasis. In order to metastasize, cells migrate and invade the surrounding areas (invasion) and enter the blood (intravasation).