The intracellular localization of Ca2+, Ca2+-ATPase, Calmodulin, and Calbindin D-28KD have been studied in testes of the toad Leptodactylus chaquensis, using ultracytochemical and immunohistochemical techniques. The Ca2+ presences in the nucleus and into the mitochondria of the germ cells, together with the activity of Ca2+-ATPase detected in the nuclear envelope and mitochondrial crests, suggest the participation of this transporter in the storage of Ca2+. In Sertoli cells, Ca2+ deposits were also found in vesicles and lamellar bodies. Calmodulin and Calbindin D-28KD were revealed in the cytoplasm of both cell types. At the spermatozoon level, the cation deposits were located in the subacrosomal space and in the acrosomal vesicle. Ca2+-ATPase activity was observed in the acrosomal and plasma membranes of the gamete that suggests the existence of a transport system responsible for maintaining low cytoplasmic Ca2+ levels. The activity of Ca2+-ATPase and the location of Ca2+ deposits in gamete tail would be related to flagellar movement. The colocalization of Ca2+ deposits and their binding proteins in efferent duct cells would probably be associated with secretory activity. Considering that intracellular Ca2+ is present in different gonadal cells, this work would provide a better understanding of the cation importance in the testicular functions of this species.

Three-dimensional field ion microscopy is a powerful technique to analyze material at a truly atomic scale. Most previous studies have been made on pure, crystalline materials such as tungsten or iron. In this article, we study more complex materials, and we present the first images of an amorphous sample, showing the capability to visualize the compositional fluctuations compatible with theoretical medium order in a metallic glass (FeBSi), which is extremely challenging to observe directly using other microscopy techniques. The intensity of the spots of the atoms at the moment of field evaporation in a field ion micrograph can be used as a proxy for identifying the elemental identity of the imaged atoms. By exploiting the elemental identification and positioning information from field ion images, we show the capability of this technique to provide imaging of recrystallized phases in the annealed sample with a superior spatial resolution compared with atom probe tomography.

The mdx mouse is an experimental model of Duchenne muscular dystrophy, a genetic disorder characterized by progressive muscular degeneration which affects the oral cavity musculature, and promotes difficulty in swallowing. This study aimed to describe morphological, structural, and ultrastructural changes in the tongue mucosa and musculature of mdx mice. Forty six-month-old mice were divided into two groups: Control C57bl/10 (n = 20) and mdx C57bl/10mdx (n = 20). The tongue was dissected and analyzed with light microscopy, scanning electron microscopy, and transmission electron microscopy techniques. Our results showed conical and triangular filiform, fungiform, foliate, and vallate papillae, and their connective tissue cores. The epithelium layers identified were corneum, granulosum, spinosum, and basale. The mdx group had a thicker epithelium. Lamina propria was composed of reddish and greenish collagen. In mdx, collagen was present in the musculature of the tongue's body and in the muscular tissue between mucous and serous glands of the caudal region. Musculature was also characterized by a shorter length of sarcoplasmic invaginations, myocytolysis in mitochondrial groupings, and inflammatory focus. In conclusion, the tongue of 6-month-old mdx mice had morphology, structure, and ultrastructure revealed, showing higher wear of filiform papillae indirect reflex from the muscular degeneration process.

Morphological studies concerning the digestive system can further information on animal diets, thus aiding in the understanding of feeding behavior. Given the scarcity of information on sea turtle digestive system morphology, the aim of the present study was to describe the digestive tube (DT) morphology of Eretmochelys imbricata hatchlings to further understand the diet of these individuals in the wild. DT samples from 10 stillborn turtles (undefined sex) were analyzed at the macro and microscopic levels. The esophagus, stomach, small intestine (SI), and large intestine (LI) are described. Histologically, the DT is formed by four tunics, the mucosa, submucosa, muscular, and adventitia or serosa. The esophagus is lined by keratinized stratified squamous epithelium, while the remainder of the DT is lined by a simple columnar epithelium. The esophagus mucosa is marked by conical, pointed papillae. The stomach comprises three regions, the cardiac, fundic, and pyloric and is covered by neutral mucous granular cells. The intestinal mucosa presents absorptive cells with microvilli, neutral and acidic goblet cells, and mucosa-associated lymphoid tissue. The SI is significantly longer than the LI (p value = 0.006841). These morphological findings are strong indications of adaptations to a carnivorous diet in this hawksbill turtle age group.

Social wasps have a widespread Neotropical distribution and are important pollinators and biological control agents for pest insects. The foraging activity of wasps is influenced by biotic and abiotic factors that are detected by the antennal sensilla that vary according to species, sex, caste, and environmental conditions. This study compares the types and quantities of antennal sensilla with a scanning electron microscope between the nocturnal Apoica flavissima and the diurnal Polistes simillimus wasps. Six types of sensilla were found in the antennae of both species: placoid, coeloconic, basiconic-type 1, basiconic-type 2, trichoid-type 1, and trichoid-type 2. Sensilla chaetica were found only in the scape and pedicel of A. flavissima. In the nocturnal wasp, there are 19,132.27 ± 1,247.72 sensilla in the left and 17,746.46 ± 1,477.46 in the right antennae, whereas in the diurnal wasp 14,936.72 ± 1,271.69 in the left and 16,090.82 ± 1,345.3 in the right antennae. A. flavissima has a longer antennal length and number of sensilla than P. simillimus. The higher number of antennal sensilla in the nocturnal wasp is not linked with the antennal size. The association of antennal sensilla functions with ecological and behavioral factors of A. flavissima and P. simillimus are discussed.

Atom probe tomography (APT) has been established in the microscopic chemical and spatial analysis of metallic or semiconductors nanostructures. In recent years, and especially with the development of a transfer shuttle system and adapted preparation protocols, the field of frozen liquids has been opened up. Still, very limited knowledge is available about the evaporation and fragmentation behavior of frozen liquids in APT. In this work, efforts were made to extend the method toward organic and biological soft matter, which are mostly built from hydrocarbon chains, the evaporation and fragmentation behavior of simple alkane chains (n-tetradecanes). Tetradecane shows a very complex evaporation behavior whereby peaks of C1–C15 can be observed. Based on multihit events and the representation of these in correlation plots, more detailed information about the evaporation behavior and the decay of molecules into smaller fragments in the region near the tip can be studied. A variety of different dissociation tracks of larger molecules in their excited state and their subsequent decay in low-field regions, on the way to the detector, could be observed and the dissociation zone in the low-field region was calculated.

Epilepsy is a prevalent and chronic neurological disorder marked by recurring, uncontrollable seizures of the brain. Chronic or repeated seizures produce memory problems and induce damage to different brain regions. Histamine has been reported to have neuroprotective effects. Betahistine is a histamine analogue. The current research investigated the effects of convulsions on the cerebral cortex and hippocampus of adult male albino mice and assessed the possible protective effect of betahistine. Four groups of 40 adult male mice were organized: control, betahistine (10 mg/kg/day), pentylenetetrazole (PTZ) (40 mg/kg/ on alternate days), and Betahistine-PTZ group received betahistine 1 h before PTZ. PTZ induced a substantial rise in glutamate level and a considerable decrease in histamine level. Structural changes in the cerebral cortex and cornu ammonis (CA1) of the hippocampus were detected in the pattern of neuron degeneration. Some neurons were shrunken with dark nuclei, and others had faintly stained ones. Focal accumulation of neuroglial cells and ballooned nerve cells of the cerebral cortex were also detected. Cleaved caspase-3, glial fibrillary acidic protein, and ionized calcium-binding adaptor molecule 1 showed substantial increases, while synaptophysin expression was significantly reduced. Interestingly, these changes were less prominent in mice pretreated with betahistine. In conclusion, betahistine had shown neuroprotective properties against brain damage induced by convulsions.

The raw materials of “licorice root” in the commerce consist of roots and/or rhizomes (stolons) of different species of Glycyrrhiza. Licorice products and raw materials are frequently mislabeled and often have mixed, misidentified, or unidentified species and parts. This paper provides a detailed comparative analysis of the morpho-anatomies of the rhizomes and roots of five species of Glycyrrhiza, namely G. glabra, G. uralensis, G. echinata, G. inflata, and G. lepidota, by bright-field light microscopy and scanning electron microscopy. The studied species showed some similarities in their basic anatomical features due to the fact that they are phylogenetically closely related and belong to the same genus. However, differences in microscopic features such as the thickness of cork and medullary rays, pore frequency, and size of the vessels were observed. The rhizomes can readily be distinguished by the presence of a distinct pith. The roots lack a well-defined pith and instead have primary xylem in the center.

Ciprofloxacin (CPFX®) is potent fluoroquinolone but has severe side effects. Cinnamon (CIN) and chia seeds are potent antioxidants. The current work aimed to compare the effect of CIN extract and chia seeds on CPFX®-treated submandibular salivary glands (SMGs). Thirty-two male albino rats were divided into four groups: Group 1: received saline. Group 2: received CPFX®. Group 3: received CIN extract after 4 h of CPFX® administration. Group 4: received ground chia seeds after 4 h of CPFX® administration. After 10 days, histological, histochemical, and ultrastructural examinations were done. Different examinations illustrated normal features of SMG in Groups 1 and 3. Group 2 showed degenerative signs. Group 4 showed normal features in some areas. Statistical results illustrated that Group 2 had highest mean vacuolation area%. Highest mean of PAS optical density (OD) was for Group 2. Concerning mercuric bromophenol blue stain OD; Group 1 showed highest mean OD. CPFX® has the deteriorative effect on SMG structure and ultrastructure. It leads to increased levels of glycosaminoglycans (GAGs) and decreased levels of total proteins. CIN extract showed more ameliorative effect compared to chia seeds.

The smooth muscle contraction of the vas deferens has the important function of transporting sperm. Interstitial cells (ICs) play a critical role in the pacing and modulation of various smooth muscle organs by interactions with nerves and smooth muscle. Elucidating the three-dimensional (3D) architecture of ICs is important for understanding their spatial relationship on the mesoscale between ICs, smooth muscle cells (SMCs), and nerves. In this study, the 3D ultrastructure of ICs in the smooth muscle layer of murine vas deferens and the spatial relationships between ICs, nerves, and smooth muscles were observed using confocal laser scanning microscopy and focused ion beam/scanning electron microscopy. ICs have sheet-like structures as demonstrated by 3D observation using modern analytical techniques. Sheet-like ICs have two types of 3D structures, one flattened and the other curled. Multiple extracellular vesicle (EV)-like structures were frequently observed in ICs. Various spatial relations were observed in areas between ICs, nerves, and SMCs, which formed a complex 3D network with each other. These results suggest that ICs in the smooth muscle layer of murine vas deferens may have two subtypes with different sheet-like structures and may be involved in neuromuscular signal transmission via physical interaction and EVs.

Composite, helical nanostructures formed using cooperative interactions of liquid crystals and Au nanoparticles were studied using a scanning transmission electron microscopy (STEM) mode. The investigated helical assemblies exhibit long-range hierarchical order across length scales, as a result of the crystallization (freezing) directed growth mechanism of nanoparticle-coated twisted nanoribbons and their ability to form organized bundles. Here, STEM methods were used to reproduce the 3D structure of the Au nanoparticle double helix.

Atom probe tomography measurements of self-assembled monolayers of 1-octadecanethiol on platinum tips were performed and their fragmentation behavior under the influence of different laser powers was investigated. The carbon backbone evaporates in the form of small hydrocarbon fragments consisting of one to four carbon atoms, while sulfur evaporates exclusively as single ions. The carbon molecules evaporate at very low fields of 5.9 V/nm, while S requires a considerably higher evaporation field of 23.4 V/nm. With increasing laser power, a weak, but noticeable trend toward larger fragment sizes is observed. No hydrocarbon fragments containing S are detected, indicating that a strong S–Pt bond has formed. The observed surface coverage of S fits well with literature values and is higher for (111)-oriented samples than for (200).

The selection of high-quality sperms is critical to intracytoplasmic sperm injection, which accounts for 70–80% of in vitro fertilization (IVF) treatments. So far, sperm screening is usually performed manually by clinicians. However, the performance of manual screening is limited in its objectivity, consistency, and efficiency. To overcome these limitations, we have developed a fast and noninvasive three-stage method to characterize morphology of freely swimming human sperms in bright-field microscopy images using deep learning models. Specifically, we use an object detection model to identify sperm heads, a classification model to select in-focus images, and a segmentation model to extract geometry of sperm heads and vacuoles. The models achieve an F1-score of 0.951 in sperm head detection, a z-position estimation error within ±1.5 μm in in-focus image selection, and a Dice score of 0.948 in sperm head segmentation, respectively. Customized lightweight architectures are used for the models to achieve real-time analysis of 200 frames per second. Comprehensive morphological parameters are calculated from sperm head geometry extracted by image segmentation. Overall, our method provides a reliable and efficient tool to assist clinicians in selecting high-quality sperms for successful IVF. It also demonstrates the effectiveness of deep learning in real-time analysis of live bright-field microscopy images.

Diabetes mellitus is a serious disease worldwide and causes other associated diseases. In this study, we observed the effect of streptozotocin (STZ)-induced diabetes and benfluorex treatment on muscular capillary ultrastructure. Adult male rats were used as the test subjects and each individual was intraperitoneally injected with one dose of STZ (45 mg/kg) to induce diabetes. Doses (50 mg/kg) of benfluorex were given to the subjects with tap water by intragastric gavage application once daily for 21 days. At the end of day 21, muscle tissues were obtained from animals and examined under transmission electron microscopy. From the data obtained with the electron microscope, it was observed that the control group had typical continuous capillary vascular structures in their muscles, while STZ caused disruptive disorder of the muscle cells in the capillary wall of the STZ-diabetic group. Additionally, the thickening of the basement membrane around endothelial cells, loss of mitochondrial crista in the muscle cells, enlarged endothelial cells, and narrowed vessel lumen were observed in the muscle tissue. The findings of our study revealed that STZ-induced diabetes disrupted the vascular structure, while benfluorex partially improved it.

This work presents the microstructure of the cross-section of a newly developed Nb/Inconel 601 weld with particular attention paid to the continuity, morphology of the interface, and the microstructural changes within its vicinity. Both scanning (SEM) and transmission (TEM) electron microscopy techniques are excellent tools to analyze the microstructure that affects both mechanical and corrosion resistance properties of the obtained product. Grain size examination and their orientation together with the character of grain boundaries by the electron backscattered diffraction (EBSD) technique were performed followed by chemical composition determination across the interface with energy-dispersive X-ray spectroscopy (EDS) in SEM. Then, the microstructure observations of the mixed region located at the Nb/Inconel 601 interface using the TEM technique allowed its chemical and phase composition to be revealed.

In this study, atom probe tomography (APT) was used to investigate strontium-containing bioactive glass particles (BG-Sr10) and strontium-releasing bioactive glass-based scaffolds (pSrBG), both of which are attractive biomaterials with applications in critical bone damage repair. We outline the challenges and corresponding countermeasures of this nonconductive biomaterial for APT sample preparation and experiments, such as avoiding direct contact between focussed ion beam micromanipulators and the extracted cantilever to reduce damage during liftout. Using a low imaging voltage (≤3 kV) and current (≤500 pA) in the scanning electron microscope and a low acceleration voltage (≤2 kV) and current (≤200 pA) in the focussed ion beam prevents tip bending in the final stages of annular milling. To optimize the atom probe experiment, we considered five factors: total detected hits, multiple hits, the background level, the charge-state ratio, and the accuracy of the measured compositions, to explore the optimal laser pulse for BG-Sr10 bioactive glass. We show that a stage temperature of 30 K, 200–250 pJ laser pulse energy, 0.3% detection rate, and 200 kHz pulse rate are optimized experimental parameters for bioactive glass. The use of improved experimental preparation methods and optimized parameters resulted in a 90% successful yield of pSrBG samples by APT.

Micromorphological features of vegetative and reproductive organs in plants are considered important because they can be used as taxonomic characters. Asperula cankiriense B. Şahin & Sağıroğlu, commonly known as “Çankırı belumu”, belongs to the family Rubiaceae and generally occurs in gypsum steppes. This research reports for the first time the micromorphological characteristics of the vegetative and reproductive organs of A. cankiriense using light and scanning electron microscopy. Taxonomically critical diagnostic features, such as raphide crystals, presence and shape of trichomes, leaf cross-section shape, flower structure, and epidermal ornamentation, were described in detail. This study provided up-to-date information on the micromorphology of this newly described species and provided additional systematic information on A. cankiriense. New data about the species are reported here and will contribute to the knowledge about the family Rubiaceae.