Hyperspectral soft X-ray emission (SXE) and cathodoluminescence (CL) spectrometry have been used to investigate a carbonaceous-rich geological deposit to understand the crystallinity and morphology of the carbon and the associated quartz. Panchromatic CL maps show both the growth of the quartz and the evidence of recrystallization. A fitted CL map reveals the distribution of Ti4+ within the grains and shows subtle growth zoning, together with radiation halos from 238U decay. The sensitivity of the SXE spectrometer to carbon, together with the anisotropic X-ray emission from highly orientated pyrolytic graphite, has enabled the C Kα peak shape to be used to measure the crystal orientation of individual graphite regions. Mapping has revealed that most grains are predominantly of a single orientation, and a number of graphite grains have been investigated to demonstrate the application of this new SXE technique. A peak fitting approach to analyzing the SXE spectra was developed to project the C Kα 2pz and 2p(x+y) orbital components of the graphite. The shape of these two end-member components is comparable to those produced by electron density of states calculations. The angular sensitivity of the SXE spectrometer has been shown to be comparable to that of electron backscatter diffraction.

Teleosts exhibit enormous heterogeneity in brain morphology, especially in the patterns of the organization of cerebellum. The cerebellum of a red-tail shark that we analyzed was well-developed and included three main divisions: the valvula cerebelli, the corpus cerebelli, and the vestibulolateral lobe. Characteristically, the cerebellar cortex contained three well-distinct layers: an outer molecular, intermediate ganglionic, and inner granular layer. The ganglionic layer possessed irregularly arranged Purkinje cells and eurydendroid cells that extended their processes into the molecular layer. Both Purkinje cells and eurydendroid cells showed immunoreactivity for iNOS2. Moreover, astrocytes in the cerebellum showed the expression of glial fibrillary acidic protein. The most striking observation in the cerebellum of shark was the lack of deep cerebellar nuclei and a well-identified white matter. On the other hand, the gray substance in the spinal cord displays a characteristic pattern in its organization, in which the dorsal horns lie quite close together, giving the gray substance the shape of an inverted Y and possessing large neurons. Notably, the white matter possessed myelinated nerve fibers. The current study provides the first report on the organization of layers and neurons in the cerebellum and spinal cord of red-tail shark. This research will contribute to the neuroanatomy and evolutionary studies of the brain of Cyprinidae.

An Al–Cu–Li aerospace alloy has been investigated to determine the order in which corrosion at different types of sites occurs in AA2099-T83. Specifically, the sequence of galvanic attack on intermetallic (IM) particles and other sites of AA2099-T83 was determined as a function of time, in 0.1 M NaCl, through the use of scanning electron microscopy and electron backscatter diffraction characterization techniques. The earliest attack occurred at isolated grains and grain boundaries and on Li-containing dispersoids. Similarly, some constituent IM particles showed evidence of trenching in the surrounding alloy matrix. These IM particles included Al7Cu2Fe and another group of unidentified particles which displayed complete trenching within the first 10 min of exposure. Al13(Fe, Mn)4 were next most active followed by Al37Fe12Cu2 with Al6(Fe,Mn) and large TiB2 particles being the least active.

The current study aimed to characterize different stages of rodlet cells using light microscopy, immunohistochemistry, and transmission electron microscopy. Granular rodlet cells have a distinct granular cytoplasm. Transitional rodlet cells had distinct capsules, and immature granules. Mature rodlet cells were pear-shaped and had elongated granules. Ruptured rodlet cells had a granular cytoplasm. The affinity of rodlet cells for different histochemical techniques was detected. Immunohistochemical analysis of rodlet cells for stem cell markers such as CD117, CD34, proliferation marker, proliferating cell nuclear antigen (PCNA), endopeptidase activity; matrix metalloproteinase-9 (MPP-9) and the angiogenic factor; vascular endothelial growth factor (VEGF) was investigated. All stages of rodlet cells were expressed CD117. However, the ruptured stage was CD117-negative. The granular, transitional, and mature stages had strong CD34 immunoaffinity, while the ruptured rodlet cells were CD34-negative. The most potent immunoreactivity for PCNA was the granular rodlet cells. The transitional cells exhibited less immunoreactivity, while mature rodlet cells had no immunoaffinity for PCNA. All stages of rodlet cells had high enzyme activity as indicated by Acridine orange and exhibited strong MPP-9 immunoaffinity. VEGF is mostly expressed by granular, transitional, and mature rodlet cells. In conclusion, rodlet cells relatively had stemness properties, endopeptidase activity, express a proliferation marker, and angiogenic factors. We suggest a potential role of rodlet cells in immune defense.

Tomography using a focused ion beam (FIB) combined with a scanning electron microscope (SEM) is well-established for a wide range of conducting materials. However, performing FIB–SEM tomography on ion- and electron-beam-sensitive materials as well as poorly conducting soft materials remains challenging. Some common challenges include cross-sectioning artifacts, shadowing effects, and charging. Fully dense materials provide a planar cross section, whereas pores also expose subsurface areas of the planar cross-section surface. The image intensity of the subsurface areas gives rise to overlap between the grayscale intensity levels of the solid and pore areas, which complicates image processing and segmentation for three-dimensional (3D) reconstruction. To avoid the introduction of artifacts, the goal is to examine porous and poorly conducting soft materials as close as possible to their original state. This work presents a protocol for the optimization of FIB–SEM tomography parameters for porous and poorly conducting soft materials. The protocol reduces cross-sectioning artifacts, charging, and eliminates shadowing effects. In addition, it handles the subsurface and grayscale intensity overlap problems in image segmentation. The protocol was evaluated on porous polymer films which have both poor conductivity and pores. 3D reconstructions, with automated data segmentation, from three films with different porosities were successfully obtained.

The aim of this in vitro study was to present a method using confocal laser scanning microscopy for three-dimensional analysis of human dental enamel subjected to ceramic bracket debonding. The labial enamel surfaces of three upper central incisors were prepared and mounted in the form of standardized specimens. A sample repositioning protocol was established to enable surface measurement and analysis before and after bracket debonding. Observations were made of representative areas measuring 1,280 × 1,280 μm2, in the center of the enamel samples, as well as of the total topography (2,500 × 3,500 μm) of the bonding areas provided by the equipment software. Noncontact three-dimensional high-resolution image analyses revealed the capabilities of the employed technique and methodology to permit the examination of specific characteristics and alterations on the surfaces, before and after the debonding and finishing procedures. The new protocol was effective to provide qualitative and quantitative assessments of changes on the same dental surfaces at different trial times. The methodology constitutes a feasible tool for revealing the effects of debonding of ceramic brackets on sound and previously injured dental enamel surfaces.

Specific features of the immunohistochemical and ultrastructural organization of the hemopoietic head-kidney (HK) in adult Ctenopharyngodon idella (Valenciennes, 1844) were investigated using light and transmission electron microscopy. The HK of grass carp possessed all developmental stages of leucocytes and erythrocytes, as well as dendritic cells and epithelial reticular cells. The rodlet cells were expressed α-smooth muscle actin (SMA). In addition, macrophages were the most numerous cells in the HK, which aggregated into structures called melanomacrophage centers (MMCs). On contrary, the chromaffin and interrenal cells (ICs) were mixed and organized into large anastomosing cords, which lined the posterior cardinal veins of the HK, and associated with many blood capillaries. The ICs displayed the characteristic features of steroid-producing cells. Three types of chromaffin cells: adrenaline, noradrenaline, and small granule-containing cells were observed in the HK. Glial fibrillary acidic protein (GFAP)-positive sustentacular cells were marked among the chromaffin cells. Hemopoietic cells, immune cells, MMCs, rodlet cells, in addition to three types of chromaffin cells and one type of interrenal cells in the HK were correlated with the functional significance of the fish concerned.