This article reviews different methods used to perform quantitative X-ray microanalysis in the electron microscope and also demonstrates the urgency of measuring the fundamental parameters of X-ray generation for the development of accurate standardless quantitative methods. Using ratios of characteristic lines acquired on the same X-ray spectrum, it is shown that the Cliff and Lorimer KA-B factor can be used in a general correction method that is appropriate for all types of specimens and electron microscopes, providing that appropriate corrections are made for X-ray absorption, fluorescence, and indirect generation. Since the fundamental parameters appear in the KA-B factor, only the ratio of the ionization cross sections needs to be known, not their absolute values. In this regard, the measurement of ratios of the KA-B factor (or intensities at different beam energies of the same material with no change of beam spreading in the material) permits the validation for the best models to compute the ratio of ionization cross sections. It is shown, using this method, that the nonrelativistic Bethe equation, to compute ionization cross section, is very close to the equation of E. Casnati et al. (J Phys B15, 155–167, 1982) and also to the equations proposed by D. Bote and F. Salvat (Phys Rev A77, 042701, 2008) for the computation of the ratio of ionization cross sections. The method is extended to show that it could be used to determine the values of the Coster-Kronig transitions factors, an important fundamental parameter for the generation of L and M lines that is mostly known with poor accuracy. The detector efficiency can be measured with specimens where their intensities were measured with an energy dispersive spectrometer detector, the efficiency of which has been measured in an X-ray synchrotron (M. Alvisi et al., Microsc Microanal12, 406–415, 2006). The spatial resolution should always be computed when performing quantitative X-ray microanalysis and the equations of R. Gauvin (Microsc Microanal13(5), 354–357, 2007) for bulk materials and the one presented in this article for thin films should be used. The effects of X-rays generated by fast secondary electrons and by Auger electrons are reviewed, and their effect can be detrimental for the spatial resolution of materials involving low-energy X-ray lines, in certain specific conditions. Finally, quantitative X-ray microanalysis of heterogeneous materials is briefly reviewed.

Cathodoluminescence (CL) microanalysis provides high-sensitivity spectra and high-resolution images associated with impurity and structural defects at the nanoscale and from luminescent materials including minerals, semiconductor, and dielectric/insulating materials. Cathodoluminescence is the non-incandescent emission of light (photons) emitted from materials excited by an electron beam. CL photons (UV-VIS-NIR) are produced as a result of electronic transitions between the conduction and valence band and may also involve electronic transitions associated with defect levels within the band gap. Microscopic defects such as imperfections and impurities influence the optical, electrical, and mechanical properties of scientifically and technologically important materials. CL microanalysis is therefore a useful microanalytical technique for characterizing these properties with high sensitivity and spatial resolution. CL image resolution can range from sub-10 nm to microns, dependent on experimental arrangement (STEM-CL, SEM-CL, optical CL microscopy), electron beam parameters, specimen configuration, carrier diffusion, etc. The sensitivity of CL microanalysis is dependent on the specimen and, in the absence of self-absorption and competitive recombination centers, can be several magnitudes more sensitive than X-ray microanalysis. In particular, quantitative CL microanalysis of trace impurity concentrations while challenging has recently been shown to be possible for a range of minerals.

Haemal nodes are capsulated, lymphoid organs with haemal circulation. Their parenchyma is distributed in primary and secondary lymphatic nodes in the cortical and medullar zones. In these zones the lymphatic tissue may adopt a cord-like shape and it is composed by plasma cell, macrophages and B and T lymphocytes. Blood circulates through the sinuses situated underneath the capsule, around the trabeculae and in the marrow.

A one-day pre-meeting workshop was held on August 7, 2011, immediately preceding the annual Microscopy and Microanalysis (M&M) 2011 meeting in Nashville, Tennessee. The workshop was organized by the Aberration-Corrected Electron Microscopy (ACEM) Focused Interest Group, with an Organizing Committee consisting of Phil Batson, Dave Muller, Larry Allard, Paul Voyles, Miofang Chi, and Mike O'Keefe.

A spectroscopic protocol is proposed to implement confocal microfluorescence imaging to the analysis of microinhomogeneity in the nanocrystallization of the core of fibers belonging to a new kind of broadband fiber amplifier based on glass with embedded nanocrystals. Nanocrystallization, crucial for achieving an adequate light emission efficiency of transition metal ions in these materials, has to be as homogeneous as possible in the fiber to assure optical amplification. This requirement calls for a sensitive method for monitoring nanostructuring in oxide glasses. Here we show that mapping microfluorescence excited at 633 nm by a He-Ne laser may give a useful tool in this regard, thanks to quasi-resonant excitation of coordination defects typical of germanosilicate materials, such as nonbridging oxygens and charged Ge-O-Ge sites, whose fluorescence are shown to undergo spectral modifications when nanocrystals form into the glass. The method has been positively checked on prototypes of optical fibers—preventively characterized by means of scanning electron microscopy and energy dispersive spectroscopy—fabricated from preforms of Ni-doped Li2O-Na2O-Sb2O3-Ga2O3-GeO2-SiO2 glass in silica cladding and subjected to heat treatment to activate gallium oxide nanocrystal growth. The method indeed enables not only the mapping of the crystallization degree but also the identification of drawing-induced defects in the fiber cladding.

Sleep deprivation causes cognitive dysfunction in which impaired neuronal plasticity in hippocampus may underlie the molecular mechanisms of this deficiency. Considering calcium-mediated NMDA receptor subunit 1 (NMDAR1) and neuronal nitric oxide synthase (nNOS) activation plays an important role in the regulation of neuronal plasticity, the present study is aimed to determine whether total sleep deprivation (TSD) would impair calcium expression, together with injury of the neuronal plasticity in hippocampus. Adult rats subjected to TSD were processed for time-of-flight secondary ion mass spectrometry, NMDAR1 immunohistochemistry, nNOS biochemical assay, cytochrome oxidase histochemistry, and the Morris water maze learning test to detect ionic, neurochemical, bioenergetic as well as behavioral changes of neuronal plasticity, respectively. Results indicated that in normal rats, strong calcium signaling along with intense NMDAR1/nNOS expression were observed in hippocampal regions. Enhanced calcium imaging and neurochemical expressions corresponded well with strong bioenergetic activity and good performance of behavioral testing. However, following TSD, both calcium intensity and NMDAR1/nNOS expressions were significantly decreased. Behavioral testing also showed poor responses after TSD. As proper calcium expression is essential for maintaining hippocampal neuronal plasticity, impaired calcium expression would depress downstream NMDAR1-mediated nNOS activation, which might contribute to the initiation or development of TSD-related cognitive deficiency.

In January 2011, a Keystone Symposium on “Extracellular Matrix and Cardiovascular Remodeling” was held in Tahoe City, California. The purpose of this meeting was to focus on the general theme of extracellular matrix (ECM) roles in cardiovascular remodeling. The presentations and discussions stimulated by this meeting, which was attended by approximately 160 people, serve as the inspiration for this special section of Microscopy and Microanalysis.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

The Registration Desk at M&M 2012 is located on the Exhibit Level (one level down) inside the 3rd Street entrance to the Phoenix Convention Center North Building (See Map). Pick up your badge and Sunday Social Reception tickets at the Registration desk by 8:00 pm. The Sunday Social starts at 6:30 pm in the Convention Center Ballroom.

Variations of the mechanical properties of red blood cells that occur during their life span have long been an intriguing task for investigations. The research presented is based on noninvasive monitoring of red blood cells of different ages performed by scanning acoustic microscopy with magnitude and phase contrast. The characteristic signature of fixed cells from groups of three different ages fractionated according to mass density is obtained from the acoustic microscope images, with the data represented in polar graphs. The analysis of these data enables the determination of averaged values for the velocities of ultrasound propagating in the cells from the different groups ranging from (1,681 ± 16) m s−1 in the youngest to (1,986 ± 20) m s−1 in the oldest group. The determined bulk modulus varies with age from (3.04 ± 0.05) GPa to (4.34 ± 0.08) GPa. An approach to determine for an age-mixed population of red blood cells, collected from a healthy person, the age of the individual cells and the age dependence of the cell parameters including density, velocity, and attenuation of longitudinal polarized ultrasonic waves traveling in the cells is demonstrated.

A mathematical framework based on singular value decomposition is used to analyze the covariance among interatomic frequency distributions in spatial distribution maps (SDMs). Using this approach, singular vectors that capture the covariance within the SDM data are obtained. The structurally relevant singular vectors (SRSVs) are identified. Using the SRSVs, we extract information from z-SDMs that not only captures the offset between the atomic planes but also captures the covariance in the atomic structure among the neighborhood atomic planes. These refined z-SDMs classify the Δ(Δz) slices in the SDMs into structurally relevant information, noise, and aberrations. The SRSVs are used to construct refined xy-SDMs that provide enhanced structural information for three-dimensional atom probe tomography.

Aberration correction, after a 50 year incubation period of developing ideas and techniques while awaiting enabling technology, has transformed electron microscopy during the first dozen years of the 21st century. Some of the conditions that accompanied this transformation, the required complexity and its effect on the way microscopy is pursued, recent results that promise to change the field, and directions for the future are briefly described.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Automatic image processing of transmission electron microscopy images (TEM) is a utopia often pursued, considering the thousands of images necessary to ensure a high resolution 3D reconstruction of virus particles or other macromolecular machines.

The combination of integrated focused ion beam–scanning electron microscope (FIB-SEM) serial sectioning and imaging techniques with image analysis provided quantitative characterization of three-dimensional (3D) pigment dispersion in dried paint films. The focused ion beam in a FIB-SEM dual beam system enables great control in slicing paints, and the sectioning process can be synchronized with SEM imaging providing high quality serial cross-section images for 3D reconstruction. Application of Euclidean distance map and ultimate eroded points image analysis methods can provide quantitative characterization of 3D particle distribution. It is concluded that 3D measurement of binder distribution in paints is effective to characterize the order of pigment dispersion in dried paint films.

Hyperspectral cathodoluminescence imaging provides spectrally and spatially resolved information on luminescent materials within a single dataset. Pushing the technique toward its ultimate nanoscale spatial limit, while at the same time spectrally dispersing the collected light before detection, increases the challenge of generating low-noise images. This article describes aspects of the instrumentation, and in particular data treatment methods, which address this problem. The methods are demonstrated by applying them to the analysis of nanoscale defect features and fabricated nanostructures in III-nitride-based materials.

The glomerular basement membrane and its associated cells are critical elements in the renal ultrafiltration process. Traditionally the anionic charge associated with several carbohydrate moieties in the glomerular basement membrane are thought to form a charge selective barrier that restricts the transmembrane flux of anionic proteins across the glomerular basement membrane into the urinary space. The charge selective function, along with the size selective component of the basement membrane, serves to limit the efflux of plasma proteins from the capillary lumen. Heparan sulfate glycosaminoglycans are anionically charged carbohydrate structures attached to proteoglycan core proteins and have a role in establishing the charge selective function of the glomerular basement membrane. Although there are a large number of studies in the literature that support this concept, the results of several recent studies using molecular genetic approaches to minimize the anionic charge of the glomerular basement membrane would suggest that the role of heparan sulfate glycosaminoglycans in the glomerular capillary wall are still not yet entirely resolved, suggesting that this research area still requires new and novel exploration.

The ability to accurately reconstruct original spatial positions of field-evaporated ions emitted from a surface is fundamental to the success of atom probe tomography. As such, a clear understanding of the evolution of specimen shape and the resultant ions' trajectories during field evaporation plays an important role in improving reconstruction accuracy. To further this understanding, field-evaporation simulations of a bilayer specimen composed of two materials having an evaporation field difference of 20% were performed. The simulated field-evaporation patterns qualitatively compare favorably with experimental data, which provides confidence in the accuracy of specimen shapes predicted by the simulation. Correlations of known original atom positions with detector hit positions as a function of lateral detector position and evaporated depth were derived from the simulation. These correlations are contrasted with the current state-of-the-art reconstruction method thus outlining limitations of the current methodology. A pair of transformations are defined that take into account field-evaporated specimen shapes, and the resulting radial magnifications, to relate recorded ion positions in detector space to reconstructed atomic positions in specimen space. This novel process, when applied to simulated data, results in approximately a factor of 2 improvement in accuracy for reconstructions of interfaces with unequal fields (most general interfaces). This method is not constrained by the fundamental assumption of a hemispherical specimen shape.