Light microscopy is a powerful tool that allows for many types of samples to be examined in a rapid, easy, and nondestructive manner. Subsequent image analysis, however, is compromised by distortion of signal by instrument optics. Deconvolution of images prior to analysis allows for the recovery of lost information by procedures that utilize either a theoretically or experimentally calculated point spread function (PSF). Using a laser scanning confocal microscope (LSCM), we have imaged whole impact tracks of comet particles captured in silica aerogel, a low density, porous SiO2 solid, by the NASA Stardust mission. In order to understand the dynamical interactions between the particles and the aerogel, precise grain location and track volume measurement are required. We report a method for measuring an experimental PSF suitable for three-dimensional deconvolution of imaged particles in aerogel. Using fluorescent beads manufactured into Stardust flight-grade aerogel, we have applied a deconvolution technique standard in the biological sciences to confocal images of whole Stardust tracks. The incorporation of an experimentally measured PSF allows for better quantitative measurements of the size and location of single grains in aerogel and more accurate measurements of track morphology.

Here, we report reproducible and accurate measurement of crystallographic parameters using scanning transmission electron microscopy. This is made possible by removing drift and residual scan distortion. We demonstrate real-space lattice parameter measurements with <0.1% error for complex-layered chalcogenides Bi2Te3, Bi2Se3, and a Bi2Te2.7Se0.3 nanostructured alloy. Pairing the technique with atomic resolution spectroscopy, we connect local structure with chemistry and bonding. Combining these results with density functional theory, we show that the incorporation of Se into Bi2Te3 causes charge redistribution that anomalously increases the van der Waals gap between building blocks of the layered structure. The results show that atomic resolution imaging with electrons can accurately and robustly quantify crystallography at the nanoscale.

Microstructure evolution during the formation of B2–NiAl by high energy ball milling of equiatomic elemental mixtures was studied by X-ray diffractometer, scanning electron microscopy, and transmission electron microscopy (TEM). The crystallite size, lattice defects and ordering of the B2–NiAl were monitored via TEM as function of milling time. The diffusion reaction, Ni+Al→NiAl3 or/and Ni2Al3, occurred during high energy ball milling, and to a certain extent offered the stored energy for the explosive exothermic reaction, Ni+Al→B2–NiAl. The fine microstructure of newly formed B2–NiAl after 5 h milling involved high density defects, e.g. antiphase boundary, long range ordering domains, vacancies, and dislocations.

X-ray fluorescence (XRF) was used to image 40 histological cross-sections of bovine ovaries (n=19), focusing on structures including: antral follicles at different stages of growth or atresia, corpora lutea at three stages of development (II–IV), and capillaries, arterioles, and other blood vessels. This method identified three key trace elements [iron (Fe), zinc (Zn), and selenium (Se)] within the ovarian tissue which appeared to be localized to specific structures. Owing to minimal preprocessing of the ovaries, important high-resolution information regarding the spatial distribution of these elements was obtained with elemental trends and colocalizations of Fe and Zn apparent, as well as the infrequent appearance of Se surrounding the antrum of large follicles, as previously reported. The ability to use synchrotron radiation to measure trace element distributions in bovine ovaries at such high resolution and over such large areas could have a significant impact on understanding the mechanisms of ovarian development. This research is intended to form a baseline study of healthy ovaries which can later be extended to disease states, thereby improving our current understanding of infertility and endocrine diseases involving the ovary.

Two polyoxometalates (POMs), synthesized through a self-assembling method, were used in the treatment of streptozotocin (STZ)-induced diabetic rats. One of these nanocompounds [tris(vanadyl)-substituted tungsto-antimonate(III)-anions—POM1] was previously described in the literature, whereas the second [tris-butyltin-21-tungsto-9-antimonate(III)-anions—POM2], was prepared by us based on our original formula. In rats with STZ-induced diabetes treated with POMs (up to a cumulative dose of 4 mg/kg bodyweight at the end of the treatments), statistically significant reduced levels of blood glucose were measured after 3 weeks, as compared with the diabetic control groups (DCGs). Ultrastructural analysis of pancreatic β-cells (including the mean diameter of secretory vesicles and of their insulin granules) in the treated diabetic rats proved the POMs contribute to limitation of cellular degeneration triggered by STZ, as well as to the presence of increased amounts of insulin-containing vesicles as compared with the DCG. The two POMs also showed hepatoprotective properties when ultrastructural aspects of hepatocytes in the experimental groups of rats were studied. Based on our in vivo studies, we concluded that the two POMs tested achieved hypoglycemiant effects by preventing STZ-triggered apoptosis of pancreatic β-cells and stimulation of insulin synthesis.

Shape from focus (SFF) is a passive optical technique that reconstructs object shape from a sequence of image taken at different focus levels. In SFF techniques, computing focus measurement for each pixel in the image sequence, through a focus measure operator, is the fundamental step. Commonly used focus measure operators compute focus quality in Cartesian space and suffer from erroneous focus quality and lack in robustness. Thus, they provide erroneous depth maps. In this paper, we introduce a new focus measure operator that computes focus quality in log-polar transform (LPT) Properties of LPT, such as biological inspiration, data selection, and edge invariance, enable computation of better focus quality in the presence of noise. Moreover, instead of using a fixed patch of the image, we suggest the use of an adaptive window. The focus quality is assessed by computing variation in LPT. The effectiveness of the proposed technique is evaluated by conducting experiments using image sequences of different simulated and real objects. The comparative analysis shows that the proposed method is robust and effective in the presence of various types of noise.

The purpose of this study was to investigate micro-morphology of the resin-dentin inter-diffusion zone using two different single-bottle self-etching dentin adhesives with and without previous acid-etching, after in vitro mechanical loading stimuli. Extracted human third molars were sectioned to obtain dentin surfaces. Two different single-bottle self-etching dentin adhesives, Futurabond U and Experimental both from VOCO, were applied following the manufacturer’s instructions or after 37% phosphoric acid application. Resin-dentin interfaces were analyzed with dye assisted confocal microscopy evaluation (CLSM), including the calcium-chelation technique, xylenol orange (CLSM-XO). CLSM revealed that resin-dentin interfaces of unloaded specimens were deficiently resin-hybridized, in general. These samples showed a Rhodamine B-labeled hybrid complex and adhesive layer completely affected by fluorescein penetration (nanoleakage) through the porous resin-dentin interface, but thicker after PA-etching. Load cycling promoted an improved sealing of the resin-dentin interface at dentin, a decrease of the hybrid complex porosity, and an increment of dentin mineralization. Load cycled specimens treated with the XO technique produced a clearly outlined fluorescence due to consistent Ca-mineral deposits within the bonding interface and inside the dentinal tubules, especially when the experimental adhesive was applied.

This study examined bone tissue responses to Ti–6Al–4V alloy implants with a hard TiN coating applied by an original powder immersion reaction-assisted coating (PIRAC) nitriding method. Progression of implant fixation in the distal epiphysis and within the medullary cavity of the rat femur was evaluated between 3 days and 6 months postimplantation by scanning electron microscopy, oxytetracycline incorporation, and histochemistry. After 6 months, successful osseointegration was achieved in both epiphyseal and diaphyseal sites. Throughout, implant portions located within the epiphysis remained in close contact with bone trabeculae that gradually engulfed the implant forming a bone collar continuous with the trabecular network of the epiphysis. In the diaphysis, woven bone was first formed within the marrow cavity around the implant and later was replaced by a shell of compact bone around the implant. In general, higher osseointegration rates were measured for TiN-coated versus the uncoated implants, both in the epiphysis and in the diaphysis. In conclusion, our findings indicate an excellent long-term biocompatibility of TiN coatings applied by the PIRAC nitriding technique and superior osteoinductive ability in comparison with uncoated Ti–6Al–4V alloy. Such coatings can, therefore, be considered for improving the corrosion and wear resistance of titanium-based orthopedic implants.

When will a small or low-contrast feature, such as an embedded second-phase particle, be visible in a scanning transmission electron microscopy (STEM) X-ray map? This work illustrates a computationally inexpensive method to simulate X-ray maps and spectrum images (SIs), based upon the equations of X-ray generation and detection. To particularize the general procedure, an example of nanostructured ferritic alloy (NFA) containing nm-sized Y2Ti2O7 embedded precipitates in ferritic stainless steel matrix is chosen. The proposed model produces physically appearing simulated SI data sets, which can either be reduced to X-ray dot maps or analyzed via multivariate statistical analysis. Comparison to NFA X-ray maps acquired using three different STEM instruments match the generated simulations quite well, despite the large number of simplifying assumptions used. A figure of merit of electron dose multiplied by X-ray collection solid angle is proposed to compare feature detectability from one data set (simulated or experimental) to another. The proposed method can scope experiments that are feasible under specific analysis conditions on a given microscope. Future applications, such as spallation proton–neutron irradiations, core-shell nanoparticles, or dopants in polycrystalline photovoltaic solar cells, are proposed.

The aim of the present study was to isolate human mesenchymal stem cells (MSCs) from palatal connective and periodontal granulation tissues and to comparatively evaluate their properties. MSCs were isolated using the explant culture method. Adherence to plastic, specific antigen makeup, multipotent differentiation potential, functionality, and ultrastructural characteristics were investigated. The frequency of colony-forming unit fibroblasts for palatal-derived mesenchymal stem cells (pMSCs) was significantly higher than that of granulation tissue-derived mesenchymal stem cells (gtMSCs). A significantly higher population doubling time and lower migration potential were recorded for gtMSCs than for pMSCs. Both cell lines were positive for CD105, CD73, CD90, CD44, and CD49f, and negative for CD34, CD45, and HLA-DR, but the level of expression was different. MSCs from both sources were relatively uniform in their ultrastructure. Generally, both cell lines possessed a large, irregular-shaped euchromatic nucleus, and cytoplasm rich in mitochondria, lysosomes, and endoplasmic reticulum. The periphery of the plasma membrane displayed many small filopodia. MSCs from both cell lines were successfully differentiated into osteogenic, adiopogenic, and chondrogenic lineages. Both healthy and diseased tissues may be considered as valuable sources of MSCs for regenerative medicine owing to the high acceptance and fewer complications during harvesting.

The creation of micro-textured dental implant surfaces possessing a stimulating activity represents a challenge in implant dentistry; particularly, the formation of a thin, biologically active, calcium-phosphate layer on their surface could help to strengthen the bond to the surrounding bone. The aim of the present study was to characterize in terms of macrostructure, micro-topography and reactivity in simulated body fluid (SBF), the surface of titanium (Ti) implants blasted with TiO2 particles, acid etched with hydrofluoric acid, and activated with Ca and Mg-containing nanoparticles. Sandblasted and acid-etched implants were analyzed by ESEM-EDX (environmental scanning electron microscope with energy dispersive X-ray system) to study the micromorphology of the surface and to perform elemental X-ray microanalysis (microchemical analyses) and element mapping. ESEM-EDX analyses were performed at time 0 and after a 28-day soaking period in SBF Hank’s balanced salt solution (HBSS) following ISO 23317 (implants for surgery—in vitro evaluation for apatite-forming ability of implant materials). Microchemical analyses (weight % and atomic %) and element mapping were carried out to evaluate the relative element content, element distribution, and calcium/phosphorus (Ca/P) atomic ratio. Raman spectroscopy was used to assess the possible presence of impurities due to manufacturing and to investigate the phases formed upon HBSS soaking. Micro-morphological analyses showed a micro-textured, highly rough surface with microgrooves. Microchemical analyses showed compositional differences among the apical, middle, and distal thirds. The micro-Raman analyses of the as-received implant showed the presence of amorphous Ti oxide and traces of anatase, calcite, and a carbonaceous material derived from the decomposition of an organic component of lipidic nature (presumably used as lubricant). A uniform layer of Ca-poor calcium phosphates (CaPs) (Ca/P ratio <1.47) was observed after soaking in HBSS; the detection of the 961 cm−1 Raman band confirms this finding. These implants showed a micro-textured surface supporting the formation of CaPs when immersed in SBF. These properties may likely favor bone anchorage and healing by stimulation of mineralizing cells.

Data-constrained modeling is a method that enables three-dimensional distribution of mineral phases and porosity in a sample to be modeled based on micro-computed tomography scans acquired at different X-ray energies. Here we describe an alternative method for measuring porosity, synchrotron K-edge subtraction using xenon gas as a contrast agent. Results from both methods applied to the same Darai limestone sample are compared. Reasonable agreement between the two methods and with other porosity measurements is obtained. The possibility of a combination of data-constrained modeling and K-edge subtraction methods for more accurate sample characterization is discussed.