We here present two new methods for the characterization of fluorescent localization microscopy images obtained from immunostained brain tissue sections. Direct stochastic optical reconstruction microscopy images of 5-HT1A serotonin receptors and glial fibrillary acidic proteins in healthy cryopreserved brain tissues are analyzed. In detail, we here present two image processing methods for characterizing differences in receptor distribution on glial cells and their distribution on neural cells: One variant relies on skeleton extraction and adaptive thresholding, the other on k-means based discrete layer segmentation. Experimental results show that both methods can be applied for distinguishing classes of images with respect to serotonin receptor distribution. Quantification of nanoscopic changes in relative protein expression on particular cell types can be used to analyze degeneration in tissues caused by diseases or medical treatment.

To observe the fine distribution of minor aluminum and germanium dopants in the erbium-doped fiber (EDF) core of an optical amplifier, a sample thinning technique was applied for field emission electron probe microanalysis (FE-EPMA) together with wavelength-dispersive X-ray spectrometry. This technique significantly improved the spatial resolution without much degradation of the minimum detection limit for FE-EPMA. As such, this enabled us to observe the distribution of minor dopants in EDF. Moreover, we propose a very simple sample preparation to prevent electron-beam radiation damage, a problem involved with FE-EPMA of low-conductivity materials such as SiO2 glass, which is the main component of EDF.

Identification and accurate characterization of platinum-group minerals (PGMs) is usually a very cumbersome procedure due to their small grain size (typically below 10 µm) and inconspicuous appearance under reflected light. A novel strategy for finding PGMs and quantifying their composition was developed. It combines a mineral liberation analyzer (MLA), a point logging system, and electron probe microanalysis (EPMA).

As a first step, the PGMs are identified using the MLA. Grains identified as PGMs are then marked and coordinates recorded and transferred to the EPMA. Case studies illustrate that the combination of MLA, point logging, and EPMA results in the identification of a significantly higher number of PGM grains than reflected light microscopy. Analysis of PGMs by EPMA requires considerable effort due to the often significant overlaps between the X-ray spectra of almost all platinum-group and associated elements. X-ray lines suitable for quantitative analysis need to be carefully selected. As peak overlaps cannot be avoided completely, an offline overlap correction based on weight proportions has been developed. Results obtained with the procedure proposed in this study attain acceptable totals and atomic proportions, indicating that the applied corrections are appropriate.

We present results of cathodoluminescence (CL) investigations of high-quality zinc oxide (ZnO) nanorods obtained by an extremely fast hydrothermal method on a silicon substrate. A scanning electron microscopy (SEM) system equipped with CL allows direct comparison of SEM images and CL maps, taken from exactly the same areas of samples. Investigations are performed at a temperature of 5 K. An interlink between sample microstructure and emission properties is investigated. CL confirms a very high quality of ZnO nanorods produced by our method. In addition, the presence of super radiation effects in ZnO nanorod arrays is suggested.

The objects of this study are various local charters (cartas de foral, in Portuguese) granted by Dom Manuel I, King of Portugal (1495–1521), which substituted for medieval ones and were intended to achieve an administrative unification. These are luxuriously illuminated manuscripts, and our study aims at obtaining a better understanding of the gilding and silvering techniques applied to the parchments, in which the forais were written, between 1500 and 1520. The combined use of microscopy and X-ray spectroscopy analyses allowed us to identify the vestigial materials used for making the parchments, including products such as salt (NaCl), lime (CaO), pumice stone (SiO2+Al2O3), and chalk (CaCO3). Chalk was employed as a whitening agent to give the parchment its final color and opacity. Shell-gold and shell-silver mixed in with animal glue or gum binding media were directly applied on type 1 and 3 forais, while very thin gold leaves (<1 µm) were applied over lead-based tempera grounds (50–180 µm thick) in type 2 forais. Silver was always employed in its finest form without a further protective layer (thus its recursive state of corrosion), while gold was used in various alloy grades.

We have developed two integrated thermocouple (TC) crucible systems that allow precise measurement of sample temperature when using a furnace associated with an environmental scanning electron microscope (ESEM). Sample temperatures measured with these systems are precise (±5°C) and reliable. The TC crucible systems allow working with solids and liquids (silicate melts or ionic liquids), independent of the gas composition and pressure. These sample holder designs will allow end users to perform experiments at high temperature in the ESEM chamber with high precision control of the sample temperature.

Chloroplast division requires filamentation temperature-sensitive Z (FtsZ), a tubulin-like GTPase of cyanobacterial endosymbiotic origin. Plants and algae possess two distinct FtsZ protein families, FtsZ1 and FtsZ2 that co-assemble into a ring (Z-ring) at the division site. Z-ring assembly and disassembly and division site positioning is controlled by both positive and negative factors via their specific interactions with FtsZ1 and FtsZ2. Here we present the in planta analysis of Arabidopsis FtsZ1 and FtsZ2 turnover in the context of a native chloroplast division machinery. Fluorescence recovery after photobleaching analysis was conducted using fluorescently tagged FtsZ at wild-type (WT)-like levels. Rapid photobleaching, low signal-to-noise ratio, and phototropic movements of chloroplasts were overcome by (i) using progressive intervals in time-lapse imaging, (ii) analyzing epidermal rather than stromal chloroplasts, and (iii) employing image stack alignment during postprocessing. In plants of WT background, fluorescence recovery half-times averaged 117 and 325 s for FtsZ1 and FtsZ2, respectively. In plants lacking ARC3, the key negative regulator of FtsZ assembly, the turnover was threefold slower. The findings are discussed in the context of previous results conducted in a heterologous system.

The mean inner potential (MIP) and inelastic mean free path (IMFP) of undoped ZnTe are determined using a combination of off-axis electron holography and convergent beam electron diffraction. The ZnTe MIP is measured to be 13.7±0.6 V, agreeing with previously reported simulations, and the IMFP at 200 keV is determined to be 46±2 nm for a collection angle of 0.75 mrad. Dynamical effects affecting holographic phase imaging as a function of incident beam direction for several common semiconductors are systematically studied and compared using Bloch wave simulations. These simulation results emphasize the need for careful choice of specimen orientation when carrying out quantitative electron holography studies in order to avoid erroneous phase measurements.

The aim of the present research was to trace CD34+ stromal fibroblastic cells (CD34+ SFCs) in the palatal connective tissue harvested for muco-gingival surgical procedures and in granulation tissues from periodontal pockets using immunohistochemical and transmission electron microscopy. Immunohistochemical analysis targeted the presence of three antigens: CD31, α-smooth muscle actin (α-SMA), and CD34. In the palate, CD31 staining revealed a colored inner ring of the vessels representing the endothelium, α-SMA+ was located in the medial layer of the vasculature, and CD34 was intensely expressed by endothelial cells and artery adventitial cells (considered to be CD34+ SFCs). Granulation tissue showed the same pattern for CD31+ and α-SMA, but a different staining pattern for CD34. Ultrastructural examination of the palatal tissue highlighted perivascular cells with fibroblast-like characteristics and pericytes in close spatial relationship to endothelial cells. The ultrastructural evaluation of granulation tissue sections confirmed the presence of neovasculature and the inflammatory nature of this tissue. The present study traced the presence of CD34+ SFCs and of pericytes in the palatal connective tissue thus highlighting once more its intrinsic regenerative capabilities. The clinical and systemic factors triggering mobilization and influencing the fate of local CD34+SCFs and other progenitors are issues to be further investigated.

Cold spray is a thermo-mechanical process where the velocity of the sprayed particles affects the deformation, bonding, and mechanical properties of the deposited material, in the form of splats or coatings. At high strain rates, the impact stresses are converted into heat, a phenomenon known as adiabatic shear, which leads to grain re-crystallization. Grain re-crystallization and growth are shown to have a direct impact on the mechanical properties of the cold-sprayed material. The present study ties the microstructural features within the cold-sprayed Ti splats and the substrate to the bonding mechanism and mechanical properties. High-resolution electron channeling contrast imaging, electron backscatter diffraction mapping, and nanoindentation were used to correlate the microstructure to the mechanical properties distribution within the titanium cold-spray splats. The formation of nanograins was observed at the titanium splat/substrate interface and contributed to metallurgical bonding. An increase in grain re-crystallization within the splat and substrate materials was observed with pre-heating of the substrate. In the substrate material, the predominant mechanism of deformation was twinning. A good relationship was found between the hardness and distribution of the twins within the substrate and the size distribution of the re-crystallized grains within the splats.

The analysis of a Portuguese “papier-mâché” sculpture depicting Saint Anthony is presented in this case study. Several questions were addressed such as the characteristics of the support, pigments used, and artistic technique in order to establish a possible timeline for its production. Qualitative analyses of the cross-sections and of the paper support were performed by optical microscopy using reflected light. Two polychrome layers from different periods and a rag pulped support were identified on the sculpture. The use of micro X-ray fluorescence and Raman microscopy techniques enabled the differentiation of coloring materials used in both polychromies. Semi-quantitative analyses of the gilded samples were also performed by scanning electron microscopy in combination with energy-dispersive spectroscopy allowing the determination of a common Au–Ag–Cu alloy with differences in the purity of the gold. The identified coloring materials lead us to believe that the sculpture was produced in the 19th century, being overpainted in the first half of the 20th century.

Archaeological materials present unique records on natural processes allowing the study of long-term material behaviors such as structural modifications and degradation mechanisms. The present work is focused on the chemical and microstructural characterization of four prehistoric arsenical copper artifacts. These artifacts were characterized by micro-energy dispersive X-ray fluorescence spectrometry, optical microscopy, scanning electron microscopy with X-ray microanalysis, micro-X-ray diffraction and synchrotron radiation micro-X-ray diffraction. Cu3As is the expected intermetallic arsenide in arsenical copper alloys, reported in the literature as exhibiting a hexagonal crystallographic structure. However, a cubic Cu3As phase was identified by X-ray diffraction in all of our analyzed archaeological artifacts, while the hexagonal Cu3As phase was clearly identified only in the artifact with higher arsenic content. Occurrence of the cubic arsenide in these particular objects, suggests that it was precipitated due to long-term aging at room temperature, which points to the need of a redefinition of the Cu-As equilibrium phase constitution. These results highlight the importance of understanding the impact of structural aging for the assessment of original properties of archaeological arsenical copper artifacts, such as hardness or color.