How do lumens form? Two mechanisms that come readily to mind are a wrapping model, similar to the wrapping of the myelin sheath around a neuronal process, and a solid core of cells followed by apoptosis of the central cells. Another obvious mechanism that was suggested over 100 years ago is the fusion of intracellular vacuoles. Whereas several recent studies have supported this latter mechanism, it has not yet been proven. Now, the appropriate animal model (zebrafish), the modern techniques (transgenic chimeras), dyes (green fluorescent protein and monomeric red fluorescent protein) that can be linked to proteins to label vacuoles, and two-photon imaging in real time finally have provided the strongest support yet. In an article by Makoto Kamei, Brian Saunders, Kayla Bayless, Louis Dye, George Davis, and Brant Weinstein the assembly of endothelial tubes from intracellular vacuoles was observed in vitro and in vivo.

It is well known that birds frequently attract mates by a ritual display of feathers or other ornaments. The colors for these displays are either derived from pigments, such as carotenoids, or photonic crystalline arrangements near the surface of the skin, feathers, or other ornaments. Add to this the fact that many birds can perceive light in the near-ultraviolet end of the visible spectrum and the range for these visual displays is extended. In a spectrophotometric and microscopic study by Birgitta Dresp and Keith Langley, it was demonstrated that a unique arrangement of crystal-like structures near the surface of the beak horn of the King penguin (Aptenodytes patagonicus) most likely accounts for a photonic effect that probably plays a role in their mating rituals.

Proteins in biologic membranes perform a large variety of essential functions. The fact that about one third of all genes code for membrane proteins, and that the majority of drugs target these proteins, attest to that fact. However, until now, proteins have been studied under artificial conditions, such as after being crystallized, frozen, or adsorbed to a substrate. Rui Pedro Gonçalves, Guillaume Agnus, Pierre Sens, Christine Houssin, Bernard Bartenlian, and Simon Scheuring have devised a novel setup with the atomic force microscope (AFM) to allow proteins to be probed while they are in unsupported membranes. Their method is similar in principle to methods where a small area of a membrane is sampled, such as when a piece of membrane is examined by patch clamping. The difference is that instead of attaching a membrane to the end of a pipette, it is spread across a piece of perforated Si(001).

Many people dread the visit to their dentist when they have their teeth drilled, but how long has this been going on? The most ancient evidence to date was recently described by Alfredo Coppa, Luca Bondioli, Andrea Cucian, David Frayer, Catherine Jarrige, Jean-François Jarrige, Guivron Quivron, Massimo Rossi, Massimo Vidale, and Roberto Macchiarelli and microscopy provided the convincing proof. The specimens were recovered from Mehrgarh, Pakistan, an area known to be occupied by farmers as long as 9,000 years ago. They identified four females, two males, and three individuals of undetermined gender who had a total of eleven drilled permanent teeth, all from adults. No drilled teeth from children were found. Four of the teeth were from the maxilla and seven from the mandible. All the drilling had been in the first or second molars. This led Coppa et al. to conclude that the drilling was not done for decorative purposes because on these teeth the holes, with or without decorative material inserted into them, would have been hardly visible.

How do the right cells get to the right place in lymph nodes? It is known that lymphocytes known as B cells (that originate in the bone marrow) migrate to follicles within the nodes, whereas T cells (that originate in the bone marrow and migrate to the thymus gland) reside in an adjacent region known as the paracortex. By combining confocal, electron, and intravital microscopy, Marc Bajénoff, Jackson Egen, Lily Koo, Jean Pierre Laugier, Frédéric Brau, Nicolas Glaichenhaus, and Ronald Germain have demonstrated a role for the stroma of the node in directing these cells to the appropriate location. The stromal cells that are critical in the B cell follicles are follicular dendritic cells (FDCs) and in the paracortex it's the fibroblastic reticular cells (FRCs).

In the mammalian embryo, the first axis to appear is at the time of the fifth cell division when the inner cell mass (ICM) becomes visible. The localization of the ICM on one side of a cavity formed within the cluster of dividing cells marks the embryonic-abembryonic (E-Ab) axis. This name derives from the fact the most of the embryo will develop from the ICM, whereas other tissues (the placenta, etc.) will develop from the other cells. There has been a long-standing controversy as to what determines the mammalian E-Ab axis; is the information inherently in the zygote, or is it determined after several cell divisions? In an elegant series of studies whereby dividing cells were labeled using new molecular genetic tools and then carefully followed during development, Yoko Kurotaki, Kohei Hatta, Kazuki Nakao, Yo-ichi Nabeshima, and Toshihiko Fujimori have provided an answer in a mouse model.

At the August M&M-2006 meeting in Chicago, we were standing next to our poster titled A Different Kind of Microscopy: Analyzing Features with an Automated Electron Beam when an acquaintance with long experience in electron microscopy wandered by. After a glance at the poster title, he challenged: “What's different about that?” Upon hearing our summary he asked (with what we took to be an encouraging tone of voice): “Are you going to publish this?” We had enough similar reactions from others to make that seem a good idea, and this is the result.

This discussion should begin by noting that the operative word in the title is “different,” not “new.” In point of fact, the foundations for the technique were laid in the 1970s when some workers began putting scanning electron microscopes and microprobes under software control by interfacing them to the “minicomputers” that powered the computerized x-ray analyzer units then entering the market. Even prior to this, there were a few “hard-wired” image analyzers that mechanized the process of extracting information from microscope images. Thus, automated analysis of features via an electron beam instrument is hardly a new concept.

Variable pressure scanning electron microscopy (VPSEM) coupled with other non-destructive analytical methods, such as energy dispersive (EDS) and Raman spectroscopy (RS) offers new capabilities for non-invasive imaging and chemical characterization of archaeological materials. This article underlines the application of VPSEM-EDS-RS on bioarchaeological specimens of pre-Columbian mummies from the Tarapacá Valley in northern Chile. The aim of the scientific investigations is to identify nonanatomical features and to provide qualitative and quantitative information at molecular levels, complementing the morphological record from studies in physical anthropology, in an effort to understand mortuary practices in the Tarapacá Valley and the effects of the burial environment in the preservation of mummified human remains.

On April 14, 1912, the RMS Titanic collided with an iceberg and sank in the North Atlantic, approximately 400 miles southeast of Newfoundland. Despite her double-bottom construction and series of water-tight compartments, the luxurious passenger liner, deemed ‘unsinkable’ by the popular press, sank in only two hours and 40 minutes, taking 1523 lives with her. As the largest man-made moving object of her time, the construction of the RMS Titanic was a technological feat, yet her sinking comprised one of the most famous disasters of the twentieth century. A colossal tragedy, the sinking has been shrouded in mystery ever since, and has led to unending speculation concerning the details of that fateful evening.

This mission flew by Comet 81P/Wild 2 where it collected more than 10,000 particles, ranging from tens of nanometers to hundreds of microns, and carried them to Earth for a safe landing in the Utah desert on January 15, 2006. So what? The answer depends on one's interest. Technically, it was a perfect DISCOVERY-class mission from launch to soft-landing (Figure 1). Scientifically, it delivered some of the 4.6 Gyrsold particles that went into making the planets of our solar system and for the first time ever we have samples from a known comet.

Many challenges lie ahead. First, the grains must be extracted from the capture cells, and prepared for mineralogical and chemical analyses, including stable isotopes and organic phases. Second, the NASA/JSC Stardust Curatorial Facility has to prepare and keep track of each sample allocation. Third, data collected in many laboratories around the globe will need to be synchronized for publication.

In 1965, Gordon Moore predicted that the number of components in an integrated circuit would double every year. The drive for higher performance with greater economy has been a major factor in the pursuit of Moore's Law. Device scaling is expected to continue without interruption, and products manufactured using a nominal 22nm feature size should become commercially available by the year 2011. This article will detail the technology enablers that make Moore's Law possible, and the improvements in microscopy techniques required to meet the challenges that Moore's Law presents.

The Gibeon meteorite is a differentiated iron meteorite that fell in Nambia, Africa in prehistoric times, with fragments spread over an area 70 miles wide and 230 miles long. The Gibeon fall was initially discovered in 1836, and hundreds of thousands of kilograms of fragments have been recovered. These fragments represent the iron core of a meteorite that cooled and crystallized over thousands of years (Norton 2002).

The microstructure of the Gibeon meteorite, which is primarily an iron-nickel alloy, consists of two phases: kamacite, a body-centered cubic material and taenite, a face-centered cubic material that metallurgists would refer to as ferrite and austenite respectively. This material initially crystallizes as taenite, and as the temperature decreases, transforms into kamacite. This meteorite is classified as a Fine Octahedrite (Of) with an average Nickel content of approximately 7.9%

The need for more precise image information of samples coming from fields such as materials analysis, semiconductor processing, and life sciences have pushed the boundaries of charged particle microscopy. A key limitation for the microscope maker in rising to these challenges lies in the relative technical maturity of source technology. Very few changes in sources have occurred in the last generations of tools available to the microscopist, while extensive efforts have been put into reducing aberrations that blur the probe—with their concomitant complexity and cost. Such efforts, representing incremental extensions of these technologies, cannot on their own address many of the emerging needs in nanotechnology. In addition, there are many challenges in the imaging of materials such as polymers and biological specimens that cannot be solved through resolution improvements but rather require different beam sample interaction dynamics. The helium ion microscope (HIM) is a breakthrough technology suited to such challenges.

The phase-contrast imaging of atomic lattices has now become commonplace for both Transmission Electron Microscopes (TEM) and Scanning Transmission Electron Microscopes (STEMs). Recently, however, bright-field STEM images of multi-wall carbon nanotubes (MWCNTs) recorded from an ultra-high resolution (UHR) in-lens field-emission scanning electron microscope (FE-SEM) operating at 30keV have also demonstrated lattice fringe resolution. One example of such an image containing multiple examples of fringe detail is shown in figure 1. The carbon lattice fringes were analyzed and their origin confirmed by the application of the FFT algorithms in the SMART image analysis program. The resulting power spectrum after thresholding to remove background noise (Figure 2) confirms that phase detail in the image extends down to about 5 Angstroms (0.5nm) and that well defined diffraction spots corresponding to a spacing of 3.4 Angstroms (0.34nm) generated by the (002) basal plane spacing of the graphite lattice are present.

For many years, as long ago as 1872, the Amphipleura pellucida (Ap) diatom striae and punctae have been an intense object for measuring the quality of resolution of the light microscope (LM). More recent publications have also referenced Ap and the difficulty of resolving its punctae. Thus, from 1872 to 1947,much attention was directed towards Ap but without paying much attention to its detailed valve structure. This was probably due to lack of resolution of LM to reveal the details for the Ap punctae (pores). Work by Stoermer and Pankratz did TEM analysis of Ap in 1964. While their results and mine do not agree, this could be attributed to specimen preparation and the source of the diatoms. However, there is an heretofore unresolved issue about the physical differences of Ap from different geographic locations. My current work should resolve this issue. This started as a challenge about whether Ap could be selectively oriented inside up or inside down.

Microwave-assisted preparation of histological samples has been performed for decades; what began with a few pioneering researchers has now become a routine and accepted practice in many clinical and research laboratories. Reliable, reproducible microwave protocols have been developed for a variety of operations: LM and EM processing, decalcification, fixation, special stains, antigen retrieval and more. Laboratories employing microwave procedures often do so for several compelling reasons: in addition to the expected time savings (often on the scale of orders of magnitude), improved morphology, retained immunoreactivity, and the elimination of hazardous reagents are benefits typically realized as well.

Despite the increasing availability of laboratory microwaves, consumer-grade (“kitchen”) microwaves continue to be used, almost invariably due to cost considerations. (EBS has maintained since 1992 that a kitchen microwave has no place in the lab.) At any time in the US there are hundreds of microwave models to choose from: a dizzying array of sizes, wattages, options, and configurations await the shopper.

Salary Surveys continue to create considerable interest. We still get requests for our 2004 salary survey results to this day. The current survey parallels the format of the 2004 survey, asking the identical questions to facilitate comparisons. The 2007 survey was sent by email to those Microscopy Today subscribers in the United States for whom we had email addresses. Respondents completed the survey on the internet within an eight-day window in mid-May 2007. We needed 350 respondents in order to have results accurate to ± 3%; we had 674 respondents. The survey was anonymous. Thank you to all that participated.

A number of trends in pharmaceutical manufacturing are driving rapid growth in the use of scanning electron microscopes (SEM) for high-resolution imaging and analysis. At the same time, continued development of SEM technology is enhancing system performance while simultaneously reducing usability barriers that have historically limited the acceptance of these techniques in pharmaceutical applications.

Among the most significant industry trends is a concerted effort by governmental regulatory agencies to encourage improvements in pharmaceutical manufacturing processes by applying the same scientific principles and engineering techniques that have lead to successful improvements in other industries, notably automobiles and semiconductors. The exceptionally high requirements for product quality within the pharmaceutical industry have generated a regulatory regime that imposes prohibitively high costs in both time and money to gain approval for process changes, even though the changes result in substantial performance gains.

Most of the specimens for high-resolution electron microscopy have amorphous surface layers due to contamination during observation and/or damaged surface layers during specimen preparation. Moreover, many specimens are radiation sensitive, and a part of the specimen easily becomes amorphous during the observation. These amorphous materials make clear observation of crystal structure difficult. A periodic structure may be extracted by simply using a periodic mask in Fourier space. However, this kind of mask often introduces a periodic feature in addition to the crystal structure. To reduce such artifacts a Wiener filter or an average background subtraction filter has been discussed. However, these filters do not work for non-ideal crystals, such as cylindrical crystals and nano-crystals, where a translational periodicity is limited to the order of nano-meter. In this report we improve these filters by introducing new ways to estimate a contribution from the amorphous materials.

Sample throughput is a significant issue in the microscopy lab. It is possible to improve the efficiency of analysis by changing the way we operate the microscope or upgrading the analytical system. One method to increase the efficiency of the analyst is to automate as many procedures as possible, including acquisitions. However, the most effective way to increase efficiency is to reduce the time that the analyst spends acquiring data. This is fine, but statistical confidence must not be reduced by the use of shorter acquisition times and the quality of the data or shape of the peaks must not be compromised as well (Figure 1, Table 1). To maintain the same level of statistical confidence of a longer analysis, an increase of the x-ray detection rate is needed.