Microscopy and Microanalysis最新文献

The biocompatibility and resorption characteristics of β-tricalcium phosphate (β-TCP, Ca3(PO4)2) have made it a coveted alternative for bone grafts. However, the underlying mechanisms governing the biological interactions between β-tricalcium phosphate and osteoclasts remain elusive. It has been speculated that the composition at grain boundaries might vary and affect β-TCP resorption properties. Atom probe tomography (APT) offers a quantitative approach to assess the composition of the grain boundaries, and thus advance our comprehension of the biological responses within the microstructure and chemical composition at the nanoscale. The precise quantitative analysis of chemical composition remains a notable challenge in APT, primarily due to the influence of measurement conditions on compositional accuracy. In this study, we investigated the impact of laser pulse energy on the composition of β-TCP using APT, aiming for the most precise Ca:P ratio and consistent results across multiple analyses performed with different sets of analysis conditions and on two different instruments.

The levels of nicotinamide adenine dinucleotide (NADH) dehydrogenase [ubiquinone] iron-sulfur protein 2 (NDUFS2, a subunit of NADH dehydrogenase) decrease in aged tissues, and these reductions may be partly associated with age-related conditions such as Parkinson's disease. Aging leads to many mitochondrial defects, such as biogenesis disruption, dysfunction, defects in the mitochondrial membrane potential, and production of reactive oxygen species, that may be highly related to NDUFS2 expression. The relationship between NDUFS2 and postovulatory oocyte aging in pigs remains unknown. In this study, we investigated changes in NDUFS2 expression during postovulatory aging (POA). Furthermore, NDUFS2 was knocked down via dsRNA microinjection at the MII stage to evaluate the effects on mitochondrial-related processes during POA. The mRNA expression of NDUFS2 decreased significantly after 48-h aging compared with that in fresh oocytes. NDUFS2 knockdown (KD) significantly impaired the maintenance of oocyte morphology and blastocyst development of embryos after POA. The levels of PGC1α (mitochondrial biogenesis-related proteins) decreased significantly after NDUFS2 KD, while the level of GSNOR, a protein denitrosylase, was reduced by NDUFS2 KD after 48 h of aging. These data suggest that NDUFS2 is vital for maintaining the oocyte quality during POA in pigs.

Identifying clusters of solute atoms in a matrix of solvent atoms helps to understand precipitation phenomena in alloys, for example, during the age hardening of certain aluminum alloys. Atom probe tomography datasets can deliver such information, provided that appropriate cluster identification routines are available. We investigate algorithms based on the local composition of the neighborhood of solute atoms and compare them with traditional approaches based on the local solute number density, such as the maximum separation distance method. For an ideal solid solution, the pair correlation functions of the kth nearest solute atom in the coordination number representation are derived, and the percolation threshold and the size distribution of clusters are studied. A criterion for selecting optimal control parameters based on maximizing the phase separation by the degree of clustering is proposed for a two-phase system. A map of phase compositions accessible for cluster analysis is constructed. The coordination number approach reduces the influence of density variations commonly observed in atom probe tomography data. Finally, a practical cluster analysis technique applied to the early stages of aluminum alloy aging is described.

SiO2 aggregates in styrene-butadiene rubber (SBR) were observed using ptychographic X-ray computed tomography (PXCT). The rubber composites were illuminated with X-rays focused by total reflection focusing mirrors, and the ptychographic diffraction patterns were collected using a CITIUS detector in the range of -75° to +75° angle of incidence. The projection images of the rubber composites were reconstructed with a two-dimensional resolution of 76 nm, and no significant structural changes were observed during the PXCT measurements. A three-dimensional image of the rubber composite was reconstructed with an isotropic resolution of 98 nm. Segmentation of SiO2 from the SBR, based on a histogram analysis of the phase shift, revealed a fragmented network structure of interconnected SiO2 aggregates.

Heat shock proteins (HSPs) are induced in response to stressful stimuli and play an important role in cell repair and protection. This study, using immunohistochemistry, aimed to determine whether HSPs are induced in the cerebellum of rats subjected to hyperthermia during postnatal development (PND). The results showed that unlike HSP27 and HSP70, HSP60 and HSP90 were constitutively expressed in the cerebellum of rats. However, hyperthermia induced HSP27 in the white matter (WM) and HSP70 in the Bergmann glial cells, the internal granule layer (IGL), and the WM. In the WM, HSP27 induction was only observed on days PND20, PND25, and PND30, and HSP27 expression was higher on day PND30 compared with days PND20 and PND25 (p < 0.001). In the Bergmann glial cells, HSP70 induction was only observed on days PND5, PND10, and PND20, and HSP70 expression was greater on days PND5 and PND10 compared with day PND20 (p < 0.001). In the IGL and the WM, HSP70 expression was higher on days PND20, PND25, and PND30 compared with days PND5 and PND10 (p < 0.001). These findings indicate that unlike HSP60 and HSP90, HSP27 and HSP70 have different expression patterns in the cerebellum of rats after hyperthermia during PND.

Abamectin is one of the most widely used pesticides due to its strong insecticidal and anthelmintic activities. Melatonin is a neurohormone with potent antioxidant, anti-apoptotic, and anti-inflammatory effects. This study aimed to investigate the potential ameliorative effects of melatonin against abamectin-induced testicular toxicity in rats. Twenty-four rats were divided into four groups: control group (1 mL/kg/day corn oil), melatonin-treated group (10 mg/kg/day), abamectin-treated group (0.5 mg/kg/day), and melatonin plus abamectin-treated group. Test substances were administered via oral gavage once daily for 28 days. While MDA and 8-OHdG levels increased in the testicular tissue of rats treated with abamectin, SOD, CAT, GPx, and GST enzyme activities decreased significantly. While interleukin-17 levels, TNF-α, and caspase3 expression increased in the testicular tissue, acetylcholinesterase activity decreased. At the same time, serum gonadotropins (luteinizing and follicle-stimulating hormones) and testosterone levels decreased. Light microscope examinations of testicular tissues revealed severe histopathological changes, such as atrophic hyalinized seminiferous tubules, basement membrane irregularity, degeneration, spermatogenic cell loss, and necrosis. Electron microscopy examinations revealed large vacuoles in Sertoli and spermatogenic cells, swelling and vacuolization in mitochondria, lysosomal structures, and increased pyknotic nuclei. In contrast, melatonin supplementation significantly ameliorated abamectin-induced testicular toxicity in rats through antioxidant, antiapoptotic, and anti-inflammatory mechanisms.

We present two new methods of processing data from backscattered electron signals in a scanning electron microscope to image grains and subgrains. The first combines data from multiple backscattered electron images acquired at different specimen geometries to (1) better reveal grain boundaries in recrystallized microstructures and (2) distinguish between recrystallized and unrecrystallized regions in partially recrystallized microstructures. The second utilizes spherical harmonic transform indexing of electron backscatter diffraction patterns to produce high angular resolution orientation data that enable the characterization of subgrains. Subgrains are produced during high-temperature plastic deformation and have boundary misorientation angles ranging from a few degrees down to a few hundredths of a degree. We also present an algorithm to automatically segment grains from combined backscattered electron image data or grains and subgrains from high angular resolution electron backscatter diffraction data. Together, these new techniques enable rapid measurements of individual grains and subgrains from large populations.

Probe formation in scanning electron microscope (SEM) is often reduced to objective lens action modeling based on a point-spread function or Fourier transforms. In this study, we present the first complete wave optical modeling of the whole SEM column based on plane-by-plane propagation of the electron beam wavefunction without simplifying the optical system. We identify the challenges in plane-by-plane beam propagation and show how sampling limitations produce aliased results. Through a careful selection and combination of propagators, we have developed a general wave optical propagation method that is able to overcome the aliasing problem to achieve the appropriate probe widths. Using a two-step propagator, we show that it is possible to model the electron beam distribution throughout the column from the virtual source plane to the specimen plane. We also show that our results from the wave optical simulations are consistent with the geometrical theory of probe formation. Finally, as a direct application of this method, we demonstrated that the combined effect of aberrations in the condenser lens and the probe forming objective lens cannot be accurately represented using only the objective lens. Designing beam shaping experiments and studying the effect of partial coherence can be some novel applications.

Intravital microscopy has emerged as a powerful imaging tool, which allows the visualization and precise understanding of rapid physiological processes at sites of inflammation in vivo, such as vascular permeability and leukocyte migration. Leukocyte interactions with the vascular endothelium can be characterized in the living organism in the murine cremaster muscle. Here, we present a microscopy technique using an Airy Beam Light Sheet microscope that has significant advantages over our previously used confocal microscopy systems. In comparison, the light sheet microscope offers near isotropic optical resolution and faster acquisition speed, while imaging a larger field of view. With less invasive surgery we can significantly reduce side effects such as bleeding, muscle twitching, and surgical inflammation. However, the increased acquisition speed requires exceptional tissue stability to avoid imaging artefacts. Since respiratory motion is transmitted to the tissue under investigation, we have developed a relocation algorithm that removes motion artefacts from our intravital microscopy images. Using these techniques, we are now able to obtain more detailed 3D time-lapse images of the cremaster vascular microcirculation, which allow us to observe the process of leukocyte emigration into the surrounding tissue with increased temporal resolution in comparison to our previous confocal approach.

It is important to understand the carbide distribution around high-energy sites such as dislocations and grain boundaries in martensitic steels as they have a major influence on the alloy performance. The aim of this study is to characterize fine ε carbides precipitated in low-carbon lath martensitic steel using the ultrawide field-of-view (FoV) CAMECA Invizo 6000 atom probe. We demonstrate the advantages of the wide FoV and determine the optimum conditions for analysis, by comparing the results such as the background noise and the C++/C+ charge state ratio (CSR) between voltage-pulsed and laser-pulsed modes. Increasing the laser pulse energy decreased the background noise and the CSR, where 70 pJ laser pulse energy produced a comparable mass-to-charge ratio spectrum to that recorded in voltage-pulsed mode, with the bulk compositions of C, Si, and Mn closest to that measured using voltage-pulsed mode. Increasing laser pulse energies to above 300 pJ decreased the bulk carbon content, with a more diffuse distribution of carbon around the carbides. This paper outlines some of the important experimental considerations when performing quantitative study of carbide precipitation in low-carbon martensitic steels using the Invizo 6000, considerations that can also be applied to other ferrous and non-ferrous alloy systems.