SPMicros was born in the year 1966 and since then the Society has promoted 46 annual Congresses. These have been long recognized as a nursery for young scientists with a special interest in microscopy. The meetings of the Society, fostered by high level scientists like David Ferreira and Teixeira da Silva, to mention only the ones that have left us this year, were once marked by a exigent standard of quality that left a lasting mark in all of us.

Replication of chloroplast in plant cells is an essential process that requires co-assembly of the tubulin-like plastid division proteins FtsZ1 and FtsZ2 at mid-chloroplast to form a ring structure called the Z-ring. The Z-ring is stabilized via its interaction with the transmembrane protein ARC6 on the inner envelope membrane of chloroplasts. Plants lacking ARC6 are defective in plastid division and contain only one or two enlarged chloroplasts per cell with abnormal localization of FtsZ: instead of a single Z-ring, many short FtsZ filaments are distributed throughout the chloroplast. ARC6 is thought to be the anchoring point for FtsZ assemblies. To investigate the role of ARC6 in FtsZ anchoring, the mobility of green fluorescent protein–tagged FtsZ assemblies was assessed by single particle tracking in mutant plants lacking the ARC6 protein. Mean square displacement analysis showed that the mobility of FtsZ assemblies is to a large extent characterized by anomalous diffusion behavior (indicative of intermittent binding) and restricted diffusion suggesting that besides ARC6-mediated anchoring, an additional FtsZ-anchoring mechanism is present in chloroplasts.

Transmission electron microscopy and associated methods, such as single particle analysis, two-dimensional crystallography, helical reconstruction, and tomography, are highly data-intensive experimental sciences, which also have substantial variability in experimental technique. Object-oriented databases present an attractive alternative to traditional relational databases for situations where the experiments themselves are continually evolving. We present EMEN2, an easy to use object-oriented database with a highly flexible infrastructure originally targeted for transmission electron microscopy and tomography, which has been extended to be adaptable for use in virtually any experimental science. It is a pure object-oriented database designed for easy adoption in diverse laboratory environments and does not require professional database administration. It includes a full featured, dynamic web interface in addition to APIs for programmatic access. EMEN2 installations currently support roughly 800 scientists worldwide with over 1/2 million experimental records and over 20 TB of experimental data. The software is freely available with complete source.

Cantharidin, a type of terpenoid, is the blistering agent of blister beetles frequently used in traditional medicine. The isolation and anticancer activity of cantharidin from blister beetles, Mylabris cichorii has been recently reported by us. This study deals with changes in mitochondrial structure and function and understanding their significance in the underlying mechanism(s) in cantharidin-mediated antitumor effects in Dalton's lymphoma (DL) bearing mice. Cantharidin treatment caused the appearance of abnormal mitochondrial features which included roundish mitochondria with thickened membranes, irregularity in cristae, and appearance of small and large size vacuoles in mitochondria of DL cells. Cantharidin treatment resulted in a decrease in mitochondrial reduced glutathione, succinate dehydrogenase activity, mitochondrial membrane potential, and induced apoptosis and necrosis in DL cells. The decrease/release of mitochondrial cytochrome c were also observed after cantharidin treatment. Flow cytometry-based cell cycle analysis showed a time-dependent accumulation of the sub-G0 population of DL cells, thus, confirming the involvement of apoptosis in tumor cells in cantharidin-mediated antitumor activity. These finding signify that the apoptosis induced by cantharidin in DL cells should involve mitochondrial-dependent pathways. It is suggested that these cantharidin-mediated changes in mitochondria may play a crucial role in its antitumor activity.

Nonconductive specimens for scanning electron microscopy or X-ray microanalysis are coated with conductive carbon in order to reduce charging. But carbon film absorbs X-ray fluxes causing errors in measuring chemical composition. Especially when the carbon content is measured, carbon coating not only blocks X-rays but also becomes a source of carbon X-rays. It is thus necessary to determine how much errors are induced by carbon coating, and how thick coating is allowed for the accurate measurement. In this study, quantitative analysis of carbon on silicon carbide with carbon coating films was attempted by electron probe microanalyzer. It was found that measured carbon content increased in a nonlinear manner up to 40% with a film thickness, whereas silicon content decreased slightly. Carbon X-ray intensity was determined by computer simulation, which increased in a linear manner with the thickness. The discrepancy was due to a nucleation and growth of islands and thus a change of density with a thickening of coating film.

On behalf of the Microscopy Society of America, the Microanalysis Society, and the International Metallographic Society, we welcome you to the Microscopy & Microanalysis 2013 meeting and its associated Exhibition in Indianapolis, IN. Once again this year, the meeting promises to be the premier opportunity in our field to learn, to connect, and to experience all the latest innovations in the world of microscopy and microanalysis.

Porous silicon (PS) has been prepared using a microwave-assisted hydrofluoric acid (HF) etching method from a silicon wafer pre-implanted with 5 MeV Cu ions. The use of microbeam proton-induced X-ray emission (micro-PIXE) and microbeam Rutherford backscattering techniques reveals for the first time the capability of these techniques for studying the formation of micropores. The porous structures observed from micro-PIXE imaging results are compared to scanning electron microscope images. It was observed that the implanted copper accumulates in the same location as the pores and that at high implanted dose the pores form large-scale patterns of lines and concentric circles. This is the first work demonstrating the use of microwave-assisted HF etching in the formation of PS.

Since 2003, the annual Omaha Imaging Symposium has brought together experts in advanced biological imaging techniques for a one-day exposition of how advanced research imaging techniques foster progress in biological research. The eighth symposium in the series was held Friday, April 20, 2012, at the Harper Student Life Center of Creighton University, Omaha, Nebraska. This special section of Microscopy and Microanalysis consists of articles contributed by speakers at the symposium.

SPMicros was founded in 1966 as Sociedade Portuguesa de Microscopia and since then has promoted Congresses dedicated to microscopy each year. For many of us this was the place where we delivered our first communications and received feedback by a panel of senior scientists devoted to the promotion of scientific quality; the Society was thus crucial to the rise of more than one generation of microscopists. In 2005 the Society's designation was changed to the “Portuguese Society for Microscopy” aka SPMicros, and incorporated a strong and growing group dedicated to materials science as well as to the life sciences.

Within the central nervous system (CNS), antigen-presenting cells (APCs) play a critical role in orchestrating inflammatory responses where they present CNS-derived antigens to immune cells that are recruited from the circulation to the cerebrospinal fluid, parenchyma, and perivascular space. Available data indicate that APCs do so indirectly from outside of CNS vessels without direct access to luminal contents. Here, we applied high-resolution, dynamic intravital two-photon laser scanning microscopy to directly visualize extravascular CX3CR1+ APC behavior deep within undisrupted CNS tissues in two distinct anatomical sites under three different inflammatory stimuli. Surprisingly, we observed that CNS-resident APCs dynamically extend their cellular processes across an intact vessel wall into the vascular lumen with preservation of vessel integrity. While only a small number of APCs displayed intravascular extensions in intact, noninflamed vessels in the brain and the spinal cord, the frequency of projections increased over days in an experimental autoimmune encephalomyelitis model, whereas the number of projections remained stable compared to baseline days after tissue injury such as CNS tumor infiltration and aseptic spinal cord trauma. Our observation of this unique behavior by parenchyma CX3CR1+ cells in the CNS argues for further exploration into their functional role in antigen sampling and immune cell recruitment.

Fluorescence images present low signal-to-noise ratio (SNR), are corrupted by a type of multiplicative noise with Poisson distribution, and are affected by a time intensity decay due to photoblinking and photobleaching (PBPB) effects. The noise and the PBPB effects together make long-term biological observation very difficult. Here, a theoretical model based on the underlying quantum mechanic physics theory of the observation process associated with this type of image is presented and the common empirical weighted sum of two decaying exponentials is derived from the model. Improvement in the SNR obtained in denoising when the proposed method is used is particularly important in the last images of the sequence where temporal correlation is used to recover information that is sometimes faded and therefore useless from a visual inspection point of view. The proposed PBPB model is included in a Bayesian denoising algorithm previously proposed by the authors. Experiments with synthetic and real data are presented to validate the PBPB model and to illustrate the effectiveness of the model in denoising and reconstruction results.

This study examines the nature of highly fragile reaction products that form in low water content supercritical carbon dioxide (scCO2) using a combination of focus ion beam/scanning electron microscopy, confocal Raman spectroscopy, helium ion microscopy (HeIM), and transmission electron microscopy (TEM). HeIM images show these precipitates are fragile rosettes. Using the TEM revealed details on the interfacial structure between the newly formed surface precipitates and the underlying initial solid phases. Detailed microscopy analysis revealed that growth of the precipitates either followed a tip growth mechanism, with precipitates forming directly on the forsterite surface if the initial solid was nonporous (natural forsterite) or growth from the surface of the precipitates, where fluid was conducted through the porous (nanoforsterite) agglomerates to the growth center. Identification of the mechanism of formation of hydrated/hydroxylated magnesium carbonate compound phases is a key factor in unraveling the impact of water recycling on mineral reactivity in low water content scCO2 solutions, which has received a great deal of attention as a result of the potential for CO2 to act as an atmospheric greenhouse gas. Techniques used here to examine these fragile structures are also used to examine a wide range of fragile material surfaces.

The effect of lead and copper on apical segments of Gracilaria domingensis was examined. Over a period of 7 days, the segments were cultivated with concentrations of 5 and 10 ppm under laboratory conditions. The samples were processed for light, confocal, and electron microscopy, as well as histochemistry, to evaluate growth rates, mitochondrial activity, protein levels, chlorophyll a, phycobiliproteins, and carotenoids. After 7 days of exposure to lead and copper, growth rates were slower than control, and biomass loss was observed on copper-treated plants. Ultrastructural damage was primarily observed in the internal organization of chloroplasts and cell wall thickness. X-ray microanalysis detected lead in the cell wall, while copper was detected in both the cytoplasm and cell wall. Moreover, lead and copper exposure led to photodamage of photosynthetic pigments and, consequently, changes in photosynthesis. However, protein content and glutathione reductase activity decreased only in the copper treatments. In both treatments, decreased mitochondrial NADH dehydrogenase activity was observed. Taken together, the present study demonstrates that (1) heavy metals such as lead and copper negatively affect various morphological, physiological, and biochemical processes in G. domingensis and (2) copper is more toxic than lead in G. domingensis.

A variety of frequently encountered clinical problems lend themselves readily to investigation by analytical electron microscopy. e.g., a combination of scanning or transmission electron microscopy and energy dispersive x-ray microanalysis. The most common application is identification of xenobiotics or exogenous substances, such as localization and quantitation of inorganic particulates in lung tissues in patients with pneumoconiosis; identification of foreign materials within granulomas; and analysis of foreign bodies. Electron probe X-ray microanalysis (EPXMA) is also useful in the study of tissue reactions to various surgical implants of foreign materials. A variety of metals and other elements may be detected with energy dispersive X-ray analysis, including copper in tissues of patients with Wilson’s disease, thorium and gadolinium in patients injected with radiographic contrast agents (Figure 1), or gold in patients treated with long-term chrysotherapy. Endogenous particulates such as urinary calculi (Figure 2), gallstones, intraarticular and periarticular crystalline deposits in patients with rheumatic disease, dystrophic or metastatic calcifications, and hemosiderin may be analyzed rapidly and efficiently by means of EDX. Certain organometallic drugs such as amiodarone (iodine) or sodium stibogluconate (antimony) may also be detected in human tissues. Analytical electron microscopy has been a useful adjunct to forensic pathology for many years in diverse areas such as identification of trace evidence constituents or detection of arsenic or lead in victims with heavy metal poisoning. The detailed elucidation of anatomic, physiologic, and pathologic conditions provided by analytical electron microscopy is a useful diagnostic and investigative tool in clinical medicine; the analytical results often have diagnostic, therapeutic, and/or medicolegal implications. This imaging technology should grow in utility in the future as it is complemented by other techniques such as mass spectrometry, and laser Raman and infrared microspectroscopy.

Extended abstract of a paper presented at Microscopy and Microanalysis 2013 in Indianapolis, Indiana, USA, August 4 – August 8, 2013.

The Registration Desk at M&M 2013 is located on the Exhibit (Street) Level outside Hall D in the Indiana Convention Center (See Floor Plan/Map). Pick up your badge and Sunday Social Reception tickets at the Registration desk before 8:00 pm on Sunday. The Sunday Social starts at 6:30 pm in the Convention Center's Sagamore Ballroom.

In variable phase bright-field contrast, a bright-field image based on axial or concentric-peripheral light is optically superimposed with a phase-contrast image, so that typical details that are imminent in one or the other technique contribute to the resulting composite image. In particular, complex structured specimens consisting of high-density light absorbing details and additional low-density phase shifting components can be observed with improved clarity. As both partial images interfere with each other, fine details within thin specimens can be highlighted further by additional contrast effects based on interference. Haloing and shade-off are significantly reduced when compared with phase contrast carried out stand-alone. Our method is characterized by several technical means that are relevant for the high image quality that can be achieved: both illuminating light components associated with bright field and phase contrast are filtered at different colors and separated from each other so that they meet the specimen at different angles of incidence. The intensities of the phase-contrast- and bright-field-producing light can be selectively regulated so that the final image can be dominated by phase contrast or bright field, or be equalized. The condenser aperture diaphragm can be used for modulations of the image's appearance.

In this study, bicrystals of silver (Ag) and aluminum (Al) were used to investigate the physical spatial resolution of the electron backscatter diffraction system combining a digital image correlation method. Furthermore, the effect of the accelerating voltage and probe current was investigated on the physical spatial resolution of the lateral and longitudinal resolutions for Ag and Al, respectively. The lateral and longitudinal resolutions show high dependency on the accelerating voltage for a low atomic mass material of Al, In addition, these are almost independent of the accelerating voltage for a high atomic mass material of Ag. Moreover, the probe current does not play any role on both the lateral and longitudinal resolutions. The best lateral resolutions for Al and Ag are 40.5 and 12.1 nm at 10 kV and 1 nA, respectively. The best longitudinal resolutions of 23.2 and 80 nm were obtained at 10 kV and 1 nA for Al and Ag, respectively.