Crystalline properties of synthetic nanostructured hydroxyapatite (n-HA) were studied using high-resolution transmission electron microscopy. The focal-series-restoration technique, obtaining exit-plane wavefunction and spherical aberration-corrected images, was successfully applied for the first time in this electron-beam-susceptible material. Multislice simulations and energy dispersive X-ray spectroscopy were also employed to determine unequivocally that n-HA particles of different size preserve stoichiometric HA-like crystal structure. n-HA particles with sizes of twice the HA lattice parameter were found. These results can be used to optimize n-HA sinterization parameters to improve bioactivity.

Polyphosphates are used in drinking water to prevent the precipitation of cations such as calcium and iron. The possible negative impact of using polyphosphates is the undesirable complexation of lead that could result in elevated lead levels in consumers' tap water. Although the water industry has focused on complexation, lead polyphosphate solids such as lead pyrophosphate, Pb2P2O7, have been considered in other fields and not been shown to form in water. The ability to form lead pyrophosphate in water could have a potential impact on the strategies used to reduce lead levels in drinking water distribution systems. The objective of this work was to determine whether lead pyrophosphate could form under simulated potable drinking water conditions. Lead pyrophosphate was synthesized in water (pH 8.2, 10 mg C/L, 2.7 mg Cl2/L) after 13 days of aging. The formation of lead pyrophosphate was confirmed by X-ray diffraction and microscopy analysis. Synthesis did not require elevated temperatures or microwave assisted approaches used by past researchers. The findings suggest that lead (and possibly other metal) pyrophosphates could conceivably form in real drinking water systems, although much more work is necessary to determine the chemistry and kinetic boundaries.

Experimental low-loss electron (LLE) yields were measured as a function of loss energy for a range of elemental standards using a high-vacuum scanning electron microscope operating at 5 keV primary beam energy with losses from 0 to 1 keV. The resulting LLE yield curves were compared with Monte Carlo simulations of the LLE yield in the particular beam/sample/detector geometry employed in the experiment to investigate the possibility of modeling the LLE yield for a series of elements. Monte Carlo simulations were performed using both the Joy and Luo [Joy, D.C. & Luo, S., Scanning11(4), 176–180 (1989)] expression for the electron stopping power and recent tabulated values of Tanuma et al. [Tanuma, S. et al., Surf Interf Anal37(11), 978–988 (2005)] to assess the influence of the more recent stopping power data on the simulation results. Further simulations have been conducted to explore the influence of sample/detector geometry on the LLE signal in the case of layered samples consisting of a thin C overlayer on an elemental substrate. Experimental LLE data were collected from a range of elemental samples coated with a thin C overlayer, and comparisons with Monte Carlo simulations were used to establish the overlayer thickness.

Oocyte maturation is known to affect the chances for successful fertilization, embryonic development, establishment of pregnancy and delivery of a live, healthy, and viable offspring. Two-photon laser scanning microscopy (TPLSM) has previously been used to evaluate early embryonic development without a detectable impairment of subsequent development, but has never been applied to assess mammalian oocytes throughout in vitro maturation (IVM). Visualization of structures within live oocytes during IVM, followed by fertilization and embryo culture, may improve the understanding of oocyte maturation. To visualize structures within bovine oocytes using TPLSM, it is necessary to remove the cumulus cells that normally surround the oocyte during maturation. Repeated visualization of structures within the same oocyte is possible, if movement of the oocyte can be avoided. In this article, we describe the development of a method for repeated intravital imaging of denuded bovine oocytes using an upright TPLSM equipped with a specially constructed incubator. Oocytes were stained with Hoechst 33258, and the nuclear structures were evaluated. Oocyte fertilization rate was not affected by TPLSM exposure, but the developmental capacity of the denuded oocytes was significantly reduced. This is, to our knowledge, the first article describing repeated intravital imaging during mammalian oocyte maturation using TPLSM.

Electromagnetic response of individual boron nitride nanotubes (BNNTs) has been studied by spatially resolved electron energy loss spectroscopy (EELS). We demonstrate how dedicated EELS methods using subnanometer electron probes permit the analysis of local dielectric properties of a material on a nanometer scale. The continuum dielectric model has been used to analyze the low-loss EEL spectra recorded from these tubes. Using this model, we demonstrate the weak influence of the out-of-plane contribution to the dielectric response of BNNTs. The optical gap, which can be deduced from the measurements, is found to be equal to 5.8 ± 0.2 eV, which is close to that of the hexagonal boron nitride. This value is found to be independent of the nanotubes configuration (diameter, helicity, number of walls, and interaction between the different walls).

Genetic manipulation allows simultaneous expression of green fluorescent protein (GFP) and its derivatives with a wide variety of cellular proteins in a variety of living systems. Epifluorescent and confocal laser scanning microscopy (confocal) localization of GFP constructs within living tissue and cell cultures has become routine, but correlation of light microscopy and high resolution transmission electron microscopy (TEM) on components within identical cells has been problematic. In this study, we describe an approach that specifically localizes the position of GFP/yellow fluorescent protein (YFP) constructs within the same cultured cell imaged in the confocal and transmission electron microscopes. We present a simplified method for delivering cell cultures expressing fluorescent fusion proteins into LR White embedding media, which allows excellent GFP/YFP detection and also high-resolution imaging in the TEM. Confocal images from 0.5-μm-thick sections are overlaid atop TEM images of the same cells collected from the next serial ultrathin section. The overlay is achieved in Adobe Photoshop by making the confocal image somewhat transparent, then carefully aligning features within the confocal image over the same features visible in the TEM image. The method requires no specialized specimen preparation equipment; specimens are taken from live cultures to embedding within 8 h, and confocal transmission overlay microscopy can be completed within a few hours.

A luminescence database for minerals and materials has been complied from the literature, the aim being to create a resource that will aid in the analysis of luminescence spectral of ionic species in minerals and materials. The database is based on a range of excitation techniques and records both major and minor lines, and their activators. The luminescence techniques included in the database are cathodoluminescence, ion luminescence, and photoluminescence. When combined with other traditional X-ray measurements collected on the same region, use of the luminescence database will give additional insight into the chemistry of minerals and materials.

Cs correctors have revolutionized transmission electron microscopy (TEM) in that they substantially improve point resolution and information limit. The object information is found sharply localized within 0.1 nm, and the intensity image can therefore be interpreted reliably on an atomic scale. However, for a conventional intensity image, the object exit wave can still not be detected completely in that the phase, and hence indispensable object information is missing. Therefore, for example, atomic electric-field distributions or magnetic domain structures cannot be accessed. Off-axis electron holography offers unique possibilities to recover completely the aberration-corrected object wave with uncorrected microscopes and hence we would not need a Cs-corrected microscope for improved lateral resolution. However, the performance of holography is affected by aberrations of the recording TEM in that the signal/noise properties (“phase detection limit”) of the reconstructed wave are degraded. Therefore, we have realized off-axis electron holography with a Cs-corrected TEM. The phase detection limit improves by a factor of four. A further advantage is the possibility of fine-tuning the residual aberrations by a posteriori correction. Therefore, a combination of both methods, that is, Cs correction and off-axis electron holography, opens new perspectives for complete TEM analysis on an atomic scale.

Piezoelectric and electrostrictive responses in poled and unpoled ferroelectric and relaxor ferroelectric compositions are of importance in transducers for converting electrical to mechanical impulses and vice-versa. Sensor applications make use of the very high piezoelectric constant dijk of the converse effect, which also permit efficient conversion of electrical to mechanical response. One of the most important families of materials for piezoelectric applications is Pb(Zr,Ti)O3(PZT). The most widely studied composition of PZT lies at the boundary between the tetragonal and rhombohedral phases, known as the morphotropic phase boundary (MPB) and exhibits greatly enhanced dielectric and piezoelectric properties in bulk and thin film. In modern electronic applications, pyroelectric detectors, piezoelectric microsensors, and micromechanical pumps require the integration of PZT films into a variety of device structures. To get sufficiently large piezoelectric strains for optimization of the performance and reliability of the device, thick films in the thickness range of 5–50 μm are desired. On the other hand burying the device components within the substrate is of utmost importance for miniaturization. In comparison to traditional surface mounted components embedded ones will free surface space for a higher functionality of the device, reduce solder points and increase device reliability. Additionally, to reduce the device costs the use of flexible copper foil as substrates is of particular interest. Its high conductivity and compatibility with printed circuit boards makes copper an attractive candidate substrate for embedded application. However, depositing PZT thick films on copper is not trivial, due to the conflict between the high temperature required to sinter PZT (∼1150°C) and low melting temperature of Cu (∼1050°C), in addition to the easy oxidation of Cu. As a consequence the preparation of PZT thick films on Cu involves a complex route to decrease the ceramic sintering temperature and to control the oxygen partial pressure. So far, no successful deposition of PZT thick films on copper foils was reported.

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

Artifacts in the field evaporation behavior of small precipitates have limited the accuracy of atom probe tomography analysis of clusters and precipitates smaller than 2 nm. Here, we report on specific observations of reconstruction artifacts that were obtained in case of precipitates with radii less than 10 nm in Al alloys, focusing particularly on a shift that appears in the relative positioning of matrix and precipitate atoms. We show that this chemically dependent behavior, referred to as “chromatic aberration,” is due to the electrostatic field above the emitter and the variations in field evaporation of the elements constituting the precipitates.

Spindle movement, including spindle migration during first meiosis and spindle rotation during second meiosis, is essential for asymmetric divisions in mouse oocytes. Previous studies by others and us have shown that microfilaments are required for both spindle migration and rotation. In the present study, we aimed to further investigate the mechanism controlling spindle movement during mouse oocyte meiosis. By employing drug treatment and immunofluorescence microscopy, we showed that dynamic microtubule assembly was involved in both spindle migration and rotation. Furthermore, we found that the calcium/CaM/CaMKII pathway was important for regulating spindle rotation.

High-resolution transmission electron microscopy and electron energy loss spectroscopy (EELS) were performed on electrochemically anodized niobium and niobium oxide. Sintered anodes of Nb and NbO powders were anodized in 0.1 wt% H3PO4 at 10, 20, and 65 V to form surface Nb2O5 layers with an average anodization constant of 3.6 ± 0.2 nm/V. The anode/dielectric interfaces were continuous and the dielectric layers were amorphous except for occurrences of plate-like, orthorhombic pentoxide crystallites in both anodes formed at 65 V. Using EELS stoichiometry quantification and relative chemical shifts of the Nb M4,5 ionization edge, a suboxide transition layer at the amorphous pentoxide interface on the order of 5 nm was detected in the Nb anodes, whereas no interfacial suboxide layers were detected in the NbO anodes.

Recently, a new class of coatings based on oxynitrides has drawn much attention in the research field as well as in industrial applications, as shown by either the large numbers of recent publications on TM O N systems (TM—transition metal) such as Ti-O-N, Zr-O-N and Ta-O-N, or the development of Si O-N for opto-electronic devices. The properties of these coatings are related to the chemical composition and the structural arrangement. However, the knowledge about the structure of TM-O-N systems is very limited, especially how the structural arrangement of the non-metallic elements is in the lattices. To the best of our knowledge, only a few studies exist on the development of structural models for oxynitrides, based on XRD and/or XPS analysis, as e.g. Si-O-N and Ti-O-N, or on Mössbauer spectrometry for Fe-O-N. TEM was used scarcely for the characterization of TM-O-N coatings possibly due to the damage of the structure by the electron-irradiation as it is reported for Cr-O-N. This work is aimed at the crystallographic understanding of W-O-N sputtered films by using TEM and HR TEM techniques for complementing the information provided by XRD characterization.

Force Microscopy: Applications in Biology and Medicine. Edited by Bhanu P. Jena and J.K. Heinrich Hörber. John Wiley & Sons, Inc., Hoboken, NJ; 2006, 300 pages. ISBN 978-0-471-39628-4

The excitement of man's imagination and discovery by the scope of imaging tools, whether tele- or micro-, from 1015 to 10−15 m, ranges from the limits of the known universe to the atomic level. Light and electron microscopy opened vistas of biology and medicine with discovery of the unit of life, the cell and viruses. In the mid-1980s development of scanning probe microscopy allowed studies on live cells at the near-nanometer level. Thus, results from studies using the atomic force microscope (AFM) have given birth to a new field, “NanoCellBiology”, by providing investigators a three-dimensional view of the structure and dynamics of live cells and biomolecules at near-angstrom resolution. Applications of force microscopy to the study of physiological processes of living cells combined with biochemical and molecular biology are yielding rapid advances on how normal and diseased cells function. This exciting book contains 15 chapters by invited experts that examine cell secretory machinery and the discovery of a new cellular structure—the porosome—involved in cell secretion, properties of microbial and cell plant surfaces, cellular interactions of nanodrug delivery systems, intermolecular forces and avidity modulation of leukocyte adhesion molecules, micromechanical properties of lipid bilayers and vesicles, imaging of soft surfaces, high-speed action of biomolecules in motion, cytogenetic applications, and the study of macromolecular interactions in hemostasis and thrombosis.

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

Aberration correction leads to reduced focal depth of field in the electron microscope. This reduced depth of field can be exploited to probe specific depths within a sample, a process known as optical sectioning. An electron microscope fitted with aberration correctors for both the pre- and postspecimen optics can be used in a confocal mode that provides improved depth resolution and selectivity over optical sectioning in the scanning transmission electron microscope (STEM). In this article we survey the coherent and incoherent imaging modes that are likely to be used in scanning confocal electron microscopy (SCEM) and provide simple expressions to describe the images that result. Calculations compare the depth response of SCEM to optical sectioning in the STEM. The depth resolution in a crystalline matrix is also explored by performing a Bloch wave calculation for the SCEM geometry in which the pre- and postspecimen optics are defocused away from their confocal conditions.