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.

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.

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.

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.

Fluorescence microscopy has experienced tremendous growth over the past four decades and has facilitated major advancements in science. The classic fluorescent techniques (which include epifluorescence and confocal) allow researchers to selectively observe labeled structures with great clarity and consistency. Historically, biological scientists have been the most prolific users of fluorescence imaging. However, growing numbers of nano-materials research initiatives are now incorporating quantum dots and other fluorescent labels into their imaging protocols. This is especially true in areas where nano-materials and biological research is overlapping such as drug delivery. This nano-bio convergence, along with other advancements, has generated the need to observe highly dynamic events involving labeled and unlabeled structures in real time.

The Focused Ion Beam (FIB) instrument, originally designed for semiconductor circuit modification and repair, has found considerable utility as a tool for specimen preparation in several microscopy disciplines and for micromachining small parts. Essentially, a FIB makes very small and precise cuts into a target sample, which implies well-controlled etch rates and close tolerances of surface finish. However, redeposition can affect etch rate and final surface topographies. This work quantifies this redeposition as it modifies yields for different parameters of etching; models the influence of redeposition as applicable to all ion beam processing; and optimizes FIB processing parameters for enhanced yields.

Resource sharing has become an absolute necessity for modern scientific research because of the increasing expense and complexity of instruments, and the ever changing funding paradigm which often requires sharing of major instrumentation resource across multiple disciplines. Many universities and even large companies have set up centralized facilities to serve researchers from different departments, divisions and units to minimize expenses and maximizing usage.

Just in the materials research area alone, it is estimated that there are more than 500 mid-size facilities existing nationwide, as reported in 2005 by the Committee on Smaller Facilities of National Research Council. Resource sharing is also common in large research groups, between Principal Investigators, as well as in corporate/industrial laboratories.

If beauty is in the eye of the beholder, then microscopic photographer Andrew Paul Leonard might have a lot to explain. After all, touting the aesthetic merits of an eosinophil (picture that let alone pronounce it) hardly brings to mind the rugged splendor of Ansel Adam's “El Capitan” or the quirky languidness of Richard Avedon's “Dovima with Elephants.” Yet, one look at these starkly detailed images reveals a textured, beautiful world reminiscent of the fancy of Yellow Submarine and the graphic logic of M.C.Escher.

Imagine what a researcher 50 years ago would think of the technology available in today's modern research labs, where the ability to photograph and videotape live cell images in high resolution has spurred numerous advances in the sciences. Additionally, technology's shift from analog to digital image acquisition has enhanced scientific collaboration and analysis abilities, perhaps in ways our predecessors could never have imagined. However, the massive quantities of data that result from these image acquisition techniques, and subsequent storage requirements, have created unforeseen challenges. While we describe life science applications, the methods described in this article are equally applicable in physical sciences laboratories.

There are many techniques for 3D imaging of biological specimens such as confocal, two-photon and, wide-field fluorescence microscopy, CAT scan, MRI, and optical coherence tomography. There are also many derivatives of these techniques, each having its strengths and weaknesses. Due to the differences in resolution, depth-of-field, and field-of-view, it is often difficult to compare images from the relatively high-resolution microscopy methods to the latter lower-resolution high-volume imaging methods. Effectively making this comparison could be very powerful in relating organ or organism level information to cellular level processes.

Radiolaria are marine protozoa that have thrived in the world's oceans for millions of years. They are particularly unique among marine plankton in that they build silica skeletons, which have allowed them to be preserved in the fossil record. These skeletons are ornate and complex and often demonstrate perfect geometric form and symmetry. The complex and beautiful glass-like structures are visually interesting when examined with electron microscopy. These attributes, coupled with their availability, size, ease of mounting and preparation make them superb specimens for introducing students to the use of electron microscopy (EM).

The use of methacrylate monomers for embedding has a venerable history in microscopy. Many formulations have been developed over the years for various purposes, ranging from standard TEM observations to low-temperature embedding. Key parameters include the length of the hydrocarbon chain and the presence and kind of cross linking reagent. In the mixture of butyl and methyl methacrylate (BMM) described here, the monomers are relatively short-chained and there is no cross linker at all. This gives the polymerized material a softness that makes it rather unsuitable for TEM, but on the contrary allows the embedment to be removed after sectioning by a brief incubation in acetone. The latter property is good for immunocytochemistry because loss of the embedment means greater access for the antibody to the antigen. BMM generally preserves structure better than paraffin or glycol-methacrylate, and for this reason is a useful choice for light-level immunocytochemistry, particularly when sub-cellular resolution is desired.

Arecent discussion on the Microscopy Listserver on the subject of microtoming Si reminded many of us of this decades-old issue. Silicon is one of the easiest materials to prepare for TEM analysis with dozens of protocols available including chemical etching, electropolishing, mechanical polishing with and without ion milling, microcleaving, and the use of a focused ion beam instrument. The time involved in Si preparation can range from minutes to a few hours. Things become more complex if it is desired to prepare a Si semiconductor device with high specimen preparation spatial resolution i.e. prepping a prespecified, very small, structure.

Now and then a technologist, often times in the biological sciences, is asked to prepare a Si TEM specimen and wonders if Si can be microtomed. The results are almost always a pile of Si dust and frequently a damaged diamond knife.