Since the beginning of human civilization, people have apparently been fascinated with the reflection of light from surfaces. The creation of shiny surfaces extends from 4,000 B.C., when Neolithic farmers in ancient China polished jade objects, to the present day where modern polishing techniques are key to creating flat surfaces in the fabrication of electronic microcomponents. The question we are posing here is how does one determine when high technology was introduced in the process of polishing? Recently, Peter Lu, Nan Yao, Jenny So, George Harlow, Jianfang Lu, Genfu Wang, and Paul Chaikin offered an answer to this question. Interestingly, microscopes were used to provide the answer.

Many animals can walk up vertical surfaces or even along the ceiling. And many studies have examined how flies accomplish this impressive task. Whereas these studies have revealed the anatomy of the contact area and there have been many assumptions about the fly attachment mechanism, the main elements that contribute to the attachment force were unknown. However, Mattias Langer, J. Peter Ruppersberg, and Stanislav Gorb have successfully demonstrated that the fluid secreted from the fly's feet is a critical factor in attaching the fly to the ceiling.

It has generally been considered that cereals (such as wheat, rice, and maize) were the first crops to be cultivated by human beings. But a new study that included scanning electron microscopy, by Mordechai Kislev, Anat Hartmann, and Ofer Bar-Yosef (micrographs by Yakov Langsam) provides strong evidence that figs were the first agricultural crop in human history.

Kislev et al. recovered nine fig fruits (plus many nutlets called drupelets) from the ruins of a burned building near Jericho. The fire had carbonized the fruit which helped preserve the morphology of the specimens. The site was radiocarbon dated to 11,400 to 11,200 years ago. Microscopic analysis demonstrated that these specimens were of an edible fig that produces drupelets without embryos; that is, these fruits are sterile. Wild fertile figs have a symbiotic relationship with a wasp that plays an essential role in pollinating the fruit, but this fruit is inedible. In this study, no evidence of wasps or where a wasp could exit the fruit were found and these are features found in wild fruit. In other words, wild figs that can be pollinated and reproduce without human intervention are inedible, but edible figs require human intervention. The specimens examined in this study were of the latter variety.

In general terms, parasites that cause malaria are injected into the skin by mosquitoes. They then travel into the bloodstream and then to the liver where they invade liver cells and mature into forms called schizonts. Within each schizont, cell division produces thousands of tiny new forms called merozoites, each of which, when released into the bloodstream, is capable of infecting a red blood cell. This “traditional” pathway for malarial parasites may not be the only way these parasites travel through the body. Using some increasingly more powerful immuno-imaging tools, Rogerio Amino, Sabine Thiberge, Béatrice Martin, Susanna Celli, Spencer Shorte, Friedrich Frischknecht, and Robert Ménard have demonstrated an additional route that could have profound implications for developing effective vaccines for this major worldwide disease.

In addition to concerns about the appearance of a display, curators of art galleries are also concerned about conservation of the artwork and their authenticity. Microscopes have played a role in these latter activities since the 1930s. Various imaging techniques, including X-radiography, infrared reflectography, macrophotography, UV-fluorescence and raking light (light source at a low angle to the surface) imaging have their advantages and disadvantages. Confocal microscopy is most useful compared to the other methods for the purpose of examination of subsurface structure, but the close working distance (a few mm) makes it precarious to use on valuable masterpieces. More recently, Haida Liang, Marta Cid, Radu Cucu, George Dobre, Adrian Podoleanu, Justin Pedro, and David Sauders have demonstrated the usefulness of optical coherence tomography (OCT) for non-destructive examination of artwork. OCT, as discussed previously in this column, is more commonly used to examine biological specimens.

There are many transmission electron microscopes (TEMs) that have provided years of reliable service and good images, but they lack the positioning systems that are standard on the newest microscopes. A precise digital positioning capability is essential for many quantitative stereological studies and relocating specific features on a grid. For many of the older TEMs, retrofitting them with a digital positioning system is not an option, or at least a very expensive upgrade. Now Martin Hohmann-Marriott, William Sharp, Robert Roberson, and Robert Blankenship have described the hardware and software that offer a second life to these older microscopes.

In the field of document examination, several approaches can be taken to probe the authenticity of a piece of ancient writing. Scholars have the task of analyzing, and possibly translating, the language and content of a document, and assessing the character and quality of the script. Materials characterization plays another role. Both the document substrate and the ink can be examined to determine whether the materials and methods of creation are consistent with those known to be used during the historical period to which the document is attributed.

The evaluation and analysis of evidence using infrared, Raman, and SEM/EDX microprobe methods has advanced forensic science. Forensic science deals with the interaction of science with the law. This interaction requires that certain standards are met before scientific evidence is admitted in either a civil or criminal case. While the Court's burden of proof is different for a civil case than for a criminal case, the standards of evidence admissibility are not. Forensic scientists must defend their methods and conclusions in Court, regardless of whether they are a trace analyst identifying fibers, a drug analyst determining if the white powder found on a suspected criminal was cocaine, or a pharmaceutical researcher discovering new solid-state forms of a drug. Today, microspectroscopy is a primary technology used within all forensic science disciplines to increase the value of evidence. Modern microbeam methods are extending observations, enhancing documentation, providing additional information, aiding deductions, and testing hypotheses. This increase in value of scientific evidence is pivotal in adjudicating both civil and criminal litigation.

Orientation mapping via automated analysis of Electron Backscatter Diffraction (EBSD) patterns has developed into an established microstructural analysis tool in the electron microscopy community. From the early 1990s, when this technique became commercially available, there has been a steady increase in the data acquisition rates as shown in Figure 1. Currently, orientation mapping speeds of over 200 analyzed patterns per second have been achieved. With these types of acquisition rates now available, the strategy on how to best use EBSD and orientation mapping has also shifted. Early adopters of this technique had to allocate hours of Scanning Electron Microscope (SEM) beam time in order to collect statistically significant data. With current technology, what was collected in hours can now be obtained in minutes. The goal of this article is to introduce this high-speed orientation mapping and present results illustrating the benefits of this capability.

Over the past few years there has been a dramatic improvement in microscopy acquisition techniques, in effective imaging modalities as well as raw hardware performance. As the microscopist's available tools become more sophisticated and diverse—e.g., time-lapse, Z sectioning, multispectra, lifetime, nth harmonic, polarization, and many combinations thereof—we face a corresponding increase in complexity in the software for understanding and interpreting the resultant data. With lifetime imaging, for example, it is overwhelming to study the raw numbers; instead, an exponential curve-fitting algorithm must be applied to extract meaningful lifetime values from the mass of photon counts recorded by the instrument.

Many believe that microfluidics has the potential to do for chemistry and biology what the integrated circuit has done for electronics — integrating tremendously complex chemical and biological processes into simple easy-to-use devices that will eventually pervade our lives. While microfluidics has made great progress in the last decade — addressing many of the fundamental questions related to manipulating nanoliter volumes of chemicals and solutions — it still faces some very basic challenges as it moves out of the laboratory and into use. Perhaps most basic is the need for fast, accurate characterization of the size and shape of the microfluidic devices themselves. Conventional imaging and measurement techniques have proven adequate for initial development, but are unable to provide the speed and accuracy needed to support the continued development of microfluidic technologies.

Material scientists and engineers continue to developmaterials and structures that are ever smaller. Some of this engineering is to simply domore with less while the science of nanomaterials allows new materials to be produced with a novel range of physical and chemical properties due to the small length scales of the microstructural features of thematerials. Currently, nanoscalematerials have been produced with a diverse set of useful properties and can be found in common substances like sunscreen or technologically advanced microelectronic devices. A complete understanding of materials is based on knowledge of the processing used to produce an interesting material coupled with a full characterization of the structure that results. It is this structure/property relationship that is the basis of understanding any newmaterial developed at all length scales.

A new generation of scanning/transmission electron microscopes (S/TEM) offers a broad range of capabilities and dramatic improvements in usability, challenging the conventional perception of these techniques as needlessly difficult and expensive. Improvements in usability address both the operation of the instrument and the interpretation of results, while the cost of the analysis must be evaluated relative to the value of the information it provides—a value that has appreciated significantly with the burgeoning growth of the nanotechnology sector. A million dollar instrument is a bargain if it solves a multi-million dollar problem. Moreover, the broad range of capabilities available in current generation instruments makes careful matching of the cost of the capability with the potential value of the solution the key to sound economic choices. This article reviews the factors that typically drive TEM costs and provides examples of valuable matches in a number of applications.

It is well known that analysis using transmission electron microscopes (TEM) yields very high resolution images of thin specimens. However, the applicability of TEM analysis is not universal due to the requirement that a high internal vacuum is required. This high vacuum precludes the TEM study of living specimens or specimens in a gas or liquid environment. In order to tackle this problem, L. Marton of Universite Libre in Brussels, Belgium was the first to design an environmental cell (E-cell) in 1935 that was sealed in the tip of a TEMsample holder [1]. Marton's design included two 0.5 μm aluminum foils as upper and lower windows sandwiching a biological sample to sustain a living environment. The electron transparent windows permitted the confined biological objects to be imaged in TEMmode. Since then, environmental TEM (E-TEM) has received increasing attention from biological scientists and eventually from materials scientists as well.

The discovery that the diffracted light from a convex edge can be used to form a very high-quality, shadowcast image on any light microscope has led to a device and method, diffracted-light contrast (DLC), which will allow shadowcast imaging to be routinely performed on student/laboratory microscopes (Piekos, 1999, 2003). The surface lattice of Surirella gema was easily resolved, and micrographs comparing the subcellular details of buccal epithelial cells viewed with DLC vs. Nomarski DIC showed that, on the microscopes used, DLC was superior in both the detail it rendered and depth of field. Although the images presented revealed DLC to be an excellent technique, the full capabilities of the technique were not known at the time.

Since the earliest explorations of the microscopic world in the 17th century and continuing to the present day, microscopists have continuously faced a challenge in that there is an inherent tradeoff between the resolution of the microscope and the size of the field of view--as the resolution of the microscope is increased to resolve smaller features, the observable area of the specimen decreases proportionally. While being a long recognized characteristic of the traditional microscope design, this tradeoff is becoming a significant hindrance as optical microscopes are being used in automated systems for advanced biotech research, medical diagnostics, robotic micromanipulation, and industrial inspection.

E.H. Andrews [1] was the first to produce transmission electron micrographs that differentiated between crystalline and amorphous forms in long chain polymers. Working with thin cast films of cis-polyisoprene {natural rubber (NR)}; he examined them in the unstrained and strained states. Staining the unstrained thin films with osmium tetroxide vapors, he showed that spherulitic crystals were in light contrast whereas the darker contrasted matrix was the amorphous continuum (Figure 1).

With the recent success of the human genome project, researchers must now discover biological functions of the tens of thousands of proteins encoded in the genome. Molecular cell biology and structural biology methods are required for this work.

Modern multidimensional bioimaging methods, such as laser scanning confocal microscopy and other light microscopic techniques, have emerged as a major research methodology in this new era of informational biology. These methods all produce vast amounts of digital information and require specialized computer software to process and analyze it. Several programs intended for this purpose have become available in recent years, but microscopists around the world frequently discover them unsuitable for many of their required tasks. The reasons for this are varied, and include: extensive bugs that take too long to fix, prohibitively high prices (also for updates and bug fixes), algorithms that are undisclosed and as such unacceptable for accurate scientific work, limited sets of features dispersed into separate “modules”, complex and difficult user interfaces, and lack of flexibility in adapting to specific research situations.

The first publication on the Buckminsterfullerene carbon molecule (C60) was in 1985 by Kroto et al. [1]. Not much else happened right after that since samples of C60 were not available for further study. It was the discovery of Krätschmer and Huffman's group [2] in 1990 that really opened up the field. They found a method to produce C60 in gram quantity. This made it possible for researchers around the world to investigate C60 properties and further advanced the field of nanocarbon research. Establishing the existence of a third crystalline form of carbon is one of the most exciting fundamental discoveries in science to date.

Much effort is currently being expended in nanotechnology and other fields to build biomimetic, or nature-inspired, materials. The first step in this process is often to develop a more complete understanding of the structure and chemistry of biological systems. In this article, we will compare and contrast data collected on a common biological sample, a butterfly wing, using a variety of analytical techniques. Transmission Electron Microscopy (TEM) was used in order to perform bright field imaging of the sample cross section; Light Microscopy (LM) and Scanning Electron Microscopy (SEM) were used to provide structural information of the outer wing surface at various magnifications; Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) was used in order to image the chemical composition of the outer most surface layer; and Focused Ion Beam (FIB) techniques were used to cut (micro machine) features into the wing.