Several species of the chain-forming marine diatom genus Pseudo-nitzschia produce domoic acid (DA), a neuroexcitant amino acid, responsible for a human amnesic syndrome after consumption of toxified shellfish (ASP). Since 1995, the Portuguese monitoring program has regularly detected the presence of DA in shellfish. So far only P. australis has been implicated, and references therein. Identification of the causative species of Pseudo-nitzchia in natural samples is hampered by difficulties in identification at the light microscope level. Recent studies have shown that ultrastructural details of the wall, observed by transmission electron microscopy, allow the identification of species in good agreement with the phylogenetic and biological species concept.

The formation and the patterning of the coronary vasculature are critical to the development and pathology of the heart. Alterations in cytokine signaling and biomechanical load can alter the vascular distribution of the vessels within the heart. Changes in the physical patterning of the vasculature can have significant impacts on the relationships of the pressure-flow network and distribution of critical growth and survival factors to the tissue. Interleukin-6 (IL-6) is a pleiotropic cytokine that regulates several biological processes, including vasculogenesis. Using both immunohistological and cardioangiographic analyses, we tested the hypothesis that IL-6-loss will result in decreased vessel density, along with changes in vascular distribution. Moreover, given the impact of vascular patterning on pressure-flow and distribution mechanics, we utilized non-Euclidean geometrical fractal analysis to quantify the changes in patterning resulting from IL-6-loss. Our analyses revealed that IL-6-loss results in a decreased capillary density and increase in intercapillary distances, but does not alter vessel size or diameter. We also observed that the IL-6−/− coronary vasculature had a marked increase in fractal dimension (D value), indicating that IL-6-loss alters vascular patterning. Characterization of IL-6-loss on coronary vasculature may lend insight into the role of IL-6 in the formation and patterning of the vascular bed.

Magnetic nanocomposite materials consisting of 5.5 wt% Fe-Co alloy nanoparticles in a silica aerogel matrix, with compositions FexCo1−x of x = 0.50 and 0.67, have been synthesized by the sol-gel method. The high-resolution transmission electron microscopy images show nanoparticles consisting of single crystal grains of body-centered cubic Fe-Co alloy, with typical crystal grain diameters of approximately 4 and 7 nm for Fe0.5Co0.5 and Fe0.67Co0.33 samples, respectively. The energy dispersive X-ray (EDX) spectra summed over areas of the samples gave compositions FexCo1−x with x = 0.48 ± 0.06 and 0.68 ± 0.05. The EDX spectra obtained with the 1.5 nm probe positioned at the centers of ∼20 nanoparticles gave slightly lower concentrations of Fe, with means of ⟨x⟩ = 0.43 ± 0.01 and ⟨x⟩ = 0.64 ± 0.02, respectively. The Fe0.5Co0.5 sample was studied using electron energy loss spectroscopy (EELS), and EELS spectra summed over whole nanoparticles gave x = 0.47 ± 0.06. The EELS spectra from analysis profiles of nanoparticles show a distribution of Fe and Co that is homogeneous, i.e., x = 0.5, within a precision of at best ±0.05 in x and ±0.4 nm in position. The present microscopy results have not shown the presence of a thin layer of iron oxide, but this might be at the limit of detectability of the methods.

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

The annual Microscopy & Microanalysis meeting has proven to be the premiere meeting for scientists, technologists, and students who use microscopy or microanalysis in their research. Microscopy & Microanalysis 2009 will be no exception! The 2009 Executive Program Committee, consisting of co-chairs from each of the participating societies, has assembled a diverse and comprehensive program that will address applications in both the biological and physical sciences as well as recent and emerging trends in instrumentation and techniques. As in 2008, special emphasis has been placed on attracting fresh content and new faces. Several symposia have been designed to expand beyond the boundaries of the interdisciplinary nature of the Microscopy & Microanalysis meetings, reflecting the increasing need for collaborations between physical and biological sciences.

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

The secondary electron (SE) signal over a cleaved surface of GaAs p-i-n solar cells containing stacks of quantum wells (QWs) is analyzed by high-resolution scanning electron microscopy. The InGaAs QWs appear darker than the GaAsP barriers, which is attributed to the differences in electron affinity. This method is shown to be a powerful tool for profiling the conduction band minimum across junctions and interfaces with nanometer resolution. The intrinsic region is shown to be pinned to the Fermi level. Additional SE contrast mechanisms are discussed in relation to the dopant regions themselves as well as the AlGaAs window at the p-region. A novel method of in situ observation of the SE profile changes resulting from reverse biasing these structures shows that the built-in potential may be deduced. The obtained value of 0.7 eV is lower than the conventional bulk value due to surface effects.

This article presents a (scanning) transmission electron microscopy (TEM) study of Mn valency and its structural origin at La0.7Sr0.3MnO3/SrTiO3(0 0 1) thin film interfaces. Mn valency deviations can lead to a breakdown of ferromagnetic order and thus lower the tunneling magnetoresistance of tunnel junctions. Here, at the interface, a Mn valency reduction of 0.16 ± 0.10 compared to the film interior and an additional feature at the low energy-loss flank of the Mn-L3 line have been observed. The latter may be attributed to an elongation of the (0 0 1) plane spacing at the interface detected by geometrical phase analysis of high-resolution images. Regarding the interface geometry, high-resolution high-angle annular dark-field scanning TEM images reveal an atomically sharp interface in some regions whereas the transition appears broadened in others. This can be explained by the presence of steps. The performed measurements indicate that, among the various structure-related influences on the valency, the atomic layer termination and the local oxygen content are most important.

A comprehensive electron energy-loss spectroscopy (EELS) study of niobium (Nb) and stable Nb-oxide phases (NbO, NbO2, Nb2O5) was carried out. Part II of this work is devoted to quantitative EELS by means of experimental k-factors derived from the intensity ratio of the O-K edge and the Nb-M4,5 or Nb-M2,3 edges for all three stable Nb-oxides. The precision and accuracy of the quantification are investigated with respect to the influence of intensity-measurement energy windows, background subtraction, and sample thickness. Integration-window widths allowing optimum accuracy are determined. Owing to background-subtraction errors, the Nb-M4,5 edges rather than Nb-M2,3 are preferred for quantification. Different approaches are applied to improve the precision with regard to thickness-related errors. Thus, a precision up to ±1.5% is achieved by averaging spectra from all three reference oxides to determine a k-factor using Nb-M4,5. Using the experimental k-factor, the determination of atomic concentration ratios CNb/CO in the range of 0.4 (Nb2O5) to 1 (NbO) was found to be possible with an accuracy of 0.6% (relative deviation between measured and nominal composition), whereas ratios of calculated partial ionization cross sections lead to less accurate results.

Frangula azorica V. Grubow (Rhamnaceae), locally known as “sanguinho”, is an Azorean endemism inscribed on the IUCN Red List of Threatened Species. Morphological, chemical and biological studies on F. azorica are scarce, but preliminary studies on the bark of this species enable the identification of anthraquinone derivatives as major compounds of this plant part. This kind of compounds are present and responsible for the laxative activity of Frangulae cortex and Rhamni purshianae cortex, two well known herbal drugs of the Western Pharmacopoeias belonging to the same botanical familly (Rhamnaceae).

A variety of ways is described in which photons can be used not only for ultrafast electron microscopy but also to enormously widen the energy range of spatially-resolved electron spectroscopy. Periodic chains of femtosecond laser pulses are a particularly important and accurately timed source for single-shot imaging and diffraction as well as for several forms of pump-probe microscopy at even higher spatial resolution and sub-picosecond timing. Many exciting new fields are opened up for study by these developments. Ultrafast, single shot diffraction with intense pulses of X-rays supplemented by phase retrieval techniques may eventually offer a challenging alternative and purely photon-based route to dynamic imaging at high spatial resolution.

The development and morphology of the immature phases of Pachycrepoideus vindemmiae (Rondani, 1875) (Hymenoptera, Pteromalidae) are described from a laboratory rearing culture maintained on Ceratitis capitata (Wiedemann, 1824) (Diptera, Tephritidae) using microscopic techniques, including light and scanning electron microscopy. The surface of the chorion of the egg is granulated, and the micropyle occurs at the anterior end. The labrum of the first instar larva does not have sensilla, and the second to fourth instar larvae have setae on the head. The mature larva is characterized by the position and number of the integumental differentations (sensilla and setae). On completion of larval development, an adecticous and exarate pupa is produced. As for the adult, the mandibles of the pupae are toothed. Five larval instars are recorded, based on statistical analyses of the sizes of the larval mandibles in combination with characters such as the number of exuviae and excretion of the meconium. Developmental time from egg to adult emergence was ∼18–20 days for males and ∼21–23 days for females at 21–26°C, 55–85 relative humidity, and a 16L:8D photoperiod. The results show that the eggs and different larval instars of this parasitoid can be unambiguously identified only by scanning electron microscope.

The valence states of Mg-Al alloys are compared to those of reference materials (pure Mg and Al metals, and intermetallics). Two methods based on X-ray emission spectroscopy are proposed to determine the phases and their proportion: first, by analyzing the Al valence spectra of the Mg-rich alloys and the Mg valence spectra of the Al-rich alloys; second, by fitting with a linear combination of the reference spectra the Al spectra of the Al-rich alloys and the Mg spectra of the Mg-rich alloys. This enables us to determine that Al and Al3Mg2 are present in the 0–43.9 wt% Al composition range and Mg and Al12Mg17 are present in the 62.5–100 wt% Al composition range. In the 43.9–62.5% Al range, the alloy is single phase and an underestimation of the Al content of the alloy can be estimated from the comparison of the bandwidth of the alloy spectrum to the bandwidths of the reference spectra.

Microscopy and Microanalysis would like to thank the following sponsors for their generous support of the M&M 2009 Meeting and EXPO. (Sponsors as of May 5, 2009)

Aging is a recognized risk factor for erectile dysfunction (ED) contributing to vascular damage of penile tissue. Vascular system seems to be particularly prone to develop dysfunction along chronological aging, as supported by studies that associate aging with reduced endothelium-dependent dilation, atherosclerosis progression and angiogenic deficit in specific tissues. Cavernous tissue is particularly sensitive to atherosclerotic disease and to structural changes of the small vessels of the penis which conduces to functional impairment.

Focused ion beam nanotomography (FIB-nt) is a novel method for high resolution three-dimensional (3D) imaging. In this investigation we assess the methodological parameters related to image acquisition and data processing that are critical for obtaining reproducible microstructural results from granular materials and from complex suspensions. For this purpose three case studies are performed: (1) The precision of FIB-nt is evaluated by analyzing a reference sample with nanospheres. Due to the implementation of an automated correction procedure, drift phenomena can be removed largely from the FIB data. However, at high magnifications remaining drift components can induce problems for 3D-shape reconstructions. (2) Correct object recognition from densely packed microstructures requires specific algorithms for splitting of agglomerated particles. To establish quantitative criteria for the correct degree of splitting, a parametric study with dry portland cement is performed. It is shown that splitting with a k-value of 0.6 leads to accurate results. (3) Finally, the reproducibility of the entire cryo-FIB analysis is investigated for high pressure frozen cement suspensions. Reproducible analyses can be obtained if the magnification is adapted to the particle size. At low magnifications the small particles and their surface area are underestimated. At high magnifications representativity is questioned because local inhomogeneities can become dominant.

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

A method for the analysis of cathodoluminescence spectra is described that enables quantitative trace-element-level distributions to be mapped within minerals and materials. Cathodoluminescence intensities for a number of rare earth elements are determined by Gaussian peak fitting, and these intensities show positive correlation with independently measured concentrations down to parts per million levels. The ability to quantify cathodoluminescence spectra provides a powerful tool to determine both trace element abundances and charge state, while major elemental levels can be determined using more traditional X-ray spectrometry. To illustrate the approach, a scheelite from Kalgoorlie, Western Australia, is hyperspectrally mapped and the cathodoluminescence is calibrated against microanalyses collected using a laser ablation inductively coupled plasma mass spectrometer. Trace element maps show micron scale zoning for the rare earth elements Sm3+, Dy3+, Er3+, and Eu3+/Eu2+. The distribution of Eu2+/Eu3+ suggests that both valences of Eu have been preserved in the scheelite since its crystallization 1.63 billion years ago.

The ultrastructure of phellem cells of Quercus suber L. (cork oak) and Calotropis procera (Ait) R. Br. were analyzed using electron transmission microscopy to determine the presence or absence of plasmodesmata (PD). Different types of Q. suber cork samples were studied: one year shoots; virgin cork (first periderm), reproduction cork (traumatic periderm), and wet cork. The channel structures of PD were found in all the samples crossing adjacent cell walls through the suberin layer of the secondary wall. Calotropis phellem also showed PD crossing the cell walls of adjacent cells but in fewer numbers compared to Q. suber. In one year stems of cork oak, it was possible to follow the physiologically active PD with ribosomic accumulation next to the aperture of the channel seen in the phellogen cells to the completely obstructed channels in the dead cells that characterize the phellem tissue.