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.

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.

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.

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.

Traditionally, type specimens have not been deposited for most unicellular algae. There are few known holotypes and those that are available are often of limited use. Samples from taxa described before 1900 commonly consist of materials dried down on herbarium sheets or mica. Dried algal samples can be rehydrated, but preservation is very poor and little structure can be determined from them. Whether wet or dry, the amount of information that can be derived from such samples using conventional light microscopy is limited.

The present situation in Ecuador is not a happy one. You would think that this ought not to be the case. There are seven electron microscopes in the country and all of them are in working order (although with problems - see below), so why then are things not good. The problem begins with geography. Unlike most smaller countries, Ecuador is not dominated by a single large city. There are two major cities, locked in a continuous rivalry: Quito, the capital, is in the mountains, and Guayaquil, the larger city, is on the coast. They are about seven hours apart by road. Both cities have electron microscopes. Worse, however, is the fact that, in each city, the microscopes are in two different locations - making a total of four microscopy centers. This is too many and Ecuador can not support them all effectively.

Of all the analytical techniques in electron microscopy, X-ray energy dispersive spectroscopy (XEDS) is perhaps the most useful. It provides a quick identification of the elements and even with semiquantitative methods; a reasonable composition can be obtained. However, in the scanning electron microscopy (SEM), there are materials systems in which severe peak overlaps of heavier elements L and M lines cannot be easily deconvolved with lighter elements' K lines. In addition, without a sufficient overvoltage in the SEM, even identification of the heavier elements can be difficult. In the analytical electron microscope (AEM), there is always sufficient overvoltage to excite all of the elements' K-lines. However, all of the K-lines might not be able to be detected with commercially available instruments. This is illustrated in Fig.l where the maximum energy of the detector system might be set to 10, 20, or 40 keV.

In 1967, Bradbury published a detailed study of the optical properties of early, compound microscopes using modern photo micrographic techniques. This was followed by Bracegirdle's studies on the performance of 17th and 18th C. microscopes and later by Jones's comparative study of a Victorian microscope with an 18th C. Cuff-type microscope, signed ‘ Dollond ’. On the suggestion of the late Dr. S. Bradbury (personal communication), the present study on the assessment of a Ribright microscope includes photomicrographs of specimens in slides similar to the ones he used, i.e. a blow-fly proboscis and diatoms.

The problems being addressed by today's microscopists and microanalysts are becoming increasingly complex and data intensive. Thirty five years ago, many of us were challenged by the mere process of recording a spectral profile and then using the limited resources we had at hand to analyze the data to obtain quantitative results. Today, we have within our laptop computers the processing power of yesterday's supercomputers, however, computing power alone will not be sufficient to solve the next generation of problems.

Microscopists only dreamed about seeing atoms until 1951. The concept of atoms is believed to have originated in the writings of Democritus in 430 B.C., actually seeing images of atoms in a microscope became a fact only in 1951 with the publication by Erwin W. Müller, announcing his field ion microscope, FIM. Typical of the great scientist that he was, Müller immediately thought of ways to improve his new microscope and his goal then became achieving atomic resolution microscopy as he thought of it - the ability to not only see individual atoms, but to resolve the adjacent atoms comprising the atomic lattice of a crystal surface.

Fading of fluorochrome is a significant limitation to fluorescence microscopy. Several anti-fade agents, e.g. n-propyl gallate, are commonly used for glycerol-based mounting media (Longin et al., 1993; Ono et al., 2001). Samples mounted in glycerol must be kept at -20°C for long-term storage to prevent bacterial degradation. In contrast, fluorescent samples cleared and mounted in organic media can be stored indefinitely at room temperature.

Methyl salicylate or oil of wintergreen is an excellent clearing agent (refractive index = 1.53), which works well with a variety of fluorochromes. It has a pleasant aroma but is somewhat difficult to work with since it remains liquid after mounting. It was previously reported that shrimp embryos labeled with tubulin antibody and rhodamine-conjugated secondary antibody maintained their fluorescence after six months (Summers et al. 1993). These same samples, stained in November, 1990 and imaged by confocal microscopy for publication in Hertzler and Clark (1992), are still fluorescent after continuous storage in methyl salicylate at room temperature in the dark (Figure 1). The images of 62-cell stage shrimp embryos taken from these 1990 samples were collected with an Olympus Fluoview 300 laser scanning confocal microscope in January, 2006 in the Dept. of Biology, Central Michigan University.

As all microscopists are taught, resolution can be measured by knowing the closest distance between two points that can be discriminated in the image. There are some occasions when we want to measure something in a digital image, and the nominal resolution isn't sufficient for the task. One of these, extracting a contour from the image, was summarized in a previous research report. The principles would be valid for any filled area, such as a particle or structure from an X-ray dot map. The ground rule was set that the outermost pixel would be selected at every point on the contour. It was clear from the geometry shown in Fig. 1 that only values of 0°, 45°, 90°, 135° and 180° could be measured. Because of the stair step defect, tracing the sequence of pixel locations only gave offsets to 8 pixels, one directly below, two on the corners below, three pixels in the same location above, and one pixel on either side of the subject pixel. Therefore it was impossible to measure the true curvature of the contour.

Before SEMs were invented and when they were still of relatively poor resolution, one way to see the fine details of a sample surface was to prepare an electron-transparent replica of the sample surface and view it in a TEM. The carbon-coated surface of the sample was shadowed with a heavy metal to make a replica that mimicked the topography of the original surface, in a sample that could be viewed in the TEM.

We have found some value in this old technique; to examine second-phase particles freed from the metal matrix for EDX, diffraction, and morphology studies—while preserving the original position and distribution of the particles, historically called “extraction replication.”