The process of autophagy has been detected in the midgut epithelium of four millipede species: Julus scandinavius, Polyxenus lagurus, Archispirostreptus gigas, and Telodeinopus aoutii. It has been examined using transmission electron microscopy (TEM), which enabled differentiation of cells in the midgut epithelium, and some histochemical methods (light microscope and fluorescence microscope). While autophagy appeared in the cytoplasm of digestive, secretory, and regenerative cells in J. scandinavius and A. gigas, in the two other species, T. aoutii and P. lagurus, it was only detected in the digestive cells. Both types of macroautophagy, the selective and nonselective processes, are described using TEM. Phagophore formation appeared as the first step of autophagy. After its blind ends fusion, the autophagosomes were formed. The autophagosomes fused with lysosomes and were transformed into autolysosomes. As the final step of autophagy, the residual bodies were detected. Autophagic structures can be removed from the midgut epithelium via, e.g., atypical exocytosis. Additionally, in P. lagurus and J. scandinavius, it was observed as the neutralization of pathogens such as Rickettsia-like microorganisms. Autophagy and apoptosis ca be analyzed using TEM, while specific histochemical methods may confirm it.

The current investigation was carried out to record the final stages of the development of both middle and distal parts of quail ceca, Coturnix coturnix japonica to understand the role of ceca in digestion, immune system, and absorption. The cellular and subcellular structures, including epithelial cell height, microvillus surface area, the proportion of goblet cells, the thickness of muscle layer, and cecum diameter showed great variations during the development. An undeveloped smooth muscularis mucosa was observed for the first time on the ED5. Primordia of glands were observed on the ED7. On the ED15, the middle part exhibited two shapes of mucosal villi: tongue-shaped villi and U-shaped. The plicae and crypts of Lieberkühn were demonstrated on the hatching day. The lymphatic tissues appeared in the wall of both parts of the ceca at the 4 weeks of age. Scanning electron microscopy revealed a great difference in the mucosal surface between different regions. Telocytes were observed in-between the muscle fibers and formed a network during the post-hatching period. Because of fermentation and other bacterial or chemical processes that have been shown to occur in the ceca, this study supports two hypotheses: the cecal development is related to diet and the cecal epithelium act as a site for primary absorption of nutrients or for re-absorption of electrolytes or amino acids derived from the urine.

The aims of the present study were to: (i) evaluate the ultrastructural differences in the zona pellucida (ZP) surface between immature and mature bovine oocytes, and (ii) describe a new objective technique to measure the pores in the outer ZP. Intact cumulus–oocyte complexes (COCs) obtained from a local abattoir were immediately fixed (immature group) or submitted to in vitro maturation (IVM) at 38.5 °C for 24 h in a humidified atmosphere of 5% CO2 in air (mature group). Oocytes from both groups were morphologically evaluated via Scanning Electron Microscopy (SEM) and the images were processed in the Fiji/ImageJ software using a new objective methodology through the Trainable Weka Segmentation plugin. The average number of pores in ZP was greater (p < 0.05) in the mature group than the immature group. However, the size and circularity of pores in ZP did not differ (p > 0.05) between groups. In conclusion, it has been shown that the number of pores highlighted the main ultrastructural change in the morphology of the ZP surface of bovine oocytes during the IVM process. We have described an objective method that can be used to evaluate ultrastructural modifications of the ZP surface during oocyte maturation and early embryo development.

X-ray diffraction (XRD) and high-resolution scanning electron microscopy (SEM) have been used to characterize the silver mercury amalgam particles resting on the surface that comprise the image of five daguerreotype plates that were not gilded and that were prepared by three different contemporary daguerreotype makers. The regions of interest of the surface that were examined were overexposed, solarized, and highlight (white) areas. The XRD portion of the study shows that the two main silver mercury amalgam particles identified using the International Center for Diffraction Data PF4 + database were the Schachnerite/ζ (zeta) phase amalgam, Ag1.1Hg0.9, and the mercury silver amalgam, Ag0.65Hg0.35. On one of the daguerreotypes a third silver mercury amalgam, Moschellandsbergite, Ag2Hg3, was also identified in small concentrations. High-resolution SEM images corroborate the diffraction data and show that the crystalline nature of the silver mercury amalgam particles on all five plates to be mostly hexagonal, which would correspond to the Schachnerite/ζ (zeta) phase amalgam, and fewer rectangular solid and cubic crystals corresponding to the mercury silver amalgam.

We have used field emission scanning electron microscopy (FESEM) to study the high-resolution organization of cellulose microfibrils in onion epidermal cell walls. We frequently found that conventional “rule of thumb” conditions for imaging of biological samples did not yield high-resolution images of cellulose organization and often resulted in artifacts or distortions of cell wall structure. Here we detail our method of one-step fixation and dehydration with 100% ethanol, followed by critical point drying, ultrathin iridium (Ir) sputter coating (3 s), and FESEM imaging at a moderate accelerating voltage (10 kV) with an In-lens detector. We compare results obtained with our improved protocol with images obtained with samples processed by conventional aldehyde fixation, graded dehydration, sputter coating with Au, Au/Pd, or carbon, and low-voltage FESEM imaging. The results demonstrated that our protocol is simpler, causes little artifact, and is more suitable for high-resolution imaging of cell wall cellulose microfibrils whereas such imaging is very challenging by conventional methods.

The key role of the epididymis is contributing to sperm storage, maturation, and survival. The epididymis of camel has a unique structure called the intraepithelial gland. The present work aimed to investigate the structure of the epididymal intraepithelial gland with special references to the seasonal variation. The samples were collected from the distal part of the corpus epididymes of completely healthy mature camels (Camelus dromedarius) in the breeding and nonbreeding seasons. Tomato lectin-positive material had been demonstrated within the epididymal spermatozoa. Here, we provide the first transmission electron microscopic study for the intraepithelial gland of camel epididymis detecting the autophagy during the nonbreeding season. The autophagosomes originated from the endoplasmic reticulum, surrounding mitochondria, and located mainly next to the basement membrane. This location is probably valuable for subsequent passing of their contents into the interstitium for possible recycling. The histochemical and ultrastructural characteristics of the gland in the breeding season indicated a hyperactive secretory microenvironment enriched with the glycoprotein-producing machinery, which could be controlled by androgens. The present data suggest that the camel intraepithelial gland has a significant impact on the reproductive activity through their secretory microenvironment during the breeding season. Moreover, it recycles the unused organelles or proteins for reuse or to supply energy under stress conditions in the nonbreeding season.

The preparation of transmission electron microscopy (TEM) samples from powders is quite difficult and challenging. For powders with particles in the 1–5 μm size range, it is especially difficult to select an adequate sample preparation technique. Epoxy is commonly used to bind powder, but drawbacks, such as differential milling originating from unequal milling rates between the epoxy and powder, remain. We propose a new, simple method for preparing TEM samples. This method is especially useful for powders with particles in the 1–5 μm size range that are vulnerable to oxidation. The method uses solder as an embedding agent together with focused ion beam (FIB) milling. The powder was embedded in low-temperature solder using a conventional hot-mounting instrument. Subsequently, FIB was used to fabricate thin TEM samples via the lift-out technique. The solder proved to be more effective than epoxy in producing thin TEM samples with large areas. The problem of differential milling was mitigated, and the solder binder was more stable than epoxy under an electron beam. This methodology can be applied for preparing TEM samples from various powders that are either vulnerable to oxidation or composed of high atomic number elements.

In the development of dry self-lubricating composites, not only solid lubricant particle size and distribution are important, but also the correct selection of the solid lubricant characteristics, which should be stable, i.e. not reactive, during the whole processing. In this work, Fe+9 vol% h-BN composites were produced by uniaxial cold compaction and sintering, for which a reaction between h-BN and iron was detected after sintering at 1,150°C. The reaction phase was characterized by optical and scanning electron microscopy and identified by X-ray diffraction and energy-dispersive X-ray spectroscopy. The newly formed phase had high hardness when compared with the iron matrix. The resulting composites presented a high friction coefficient and high wear.

The gold technology in Iberia underwent an important development during the Early Iron Age (EIA) following the arrival of new technological skills from the Mediterranean region, including the use of filigree, granulation, and brazing. This work presents the microanalytical study of EIA gold jewels (22 spherical beads and four tongue pendants) recovered from three graves in southern Portugal. The set of jewelry, showing an extraordinary stylistic resemblance, was characterized by optical microscopy, micro energy-dispersive X-ray spectrometry and scanning electron microscopy with X-ray microanalysis to establish the alloy composition and production techniques. The pieces show a close technological relation, making use of artificial gold alloys with different amounts of silver (c. 40 and 25%). The decorative styles include different types of filigree (solid wires and hollow wires), while the joining techniques comprise brazing with Au-Ag-Cu solders and, probably, sintering. In spite of those technological and stylistic features, clearly pointing to a Mediterranean influence, the absence of granulation suggests an indigenous workshop where exogenous technologies/decorations were not yet fully integrated in the manufacture of such luxury items.

In order to clarify fine structures of the hypothetical meridian conduits of Chinese traditional medicine (CTM) in the skin, the present study used light and transmission electron microscopy to examine fasciae in different vertebrate species. Collagen fiber bundles and layers were arranged in a crisscross pattern, which developed into a special tissue micro-channel (TMC) network, in a manner that was analogs to the proposed skin meridian conduits. It was further revealed that tissue fluid in lateral TMC branches drained into wide longitudinal channels, which were distinctly different from lymphatic capillary. Mast cells, macrophages, and extracellular vesicles such as ectosomes and exosomes were distributed around telocytes (TCs) and their long processes (Telopodes, Tps) within the TMC. Cell junctions between TCs developed, which could enable the communication between contiguous but distant Tps. On the other hand, winding free Tps without cell junctions were also uncovered inside the TMC. Tissue fluid, cell junctions of TCs, mast cells, macrophages, and extracellular vesicles within the TMC corresponded to the circulating “气血” (“Qi-Xue”, i.e., information, message, and energy) of meridian conduits at the cytological level. These results could provide morphological evidence for the hypothesis that “meridians are the conduit for Qi-Xue circulation” in CTM.

The great potential of scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM-EDX) is in detection of unusual chemical elements included in ancient human dental calculus to verify hypotheses about life and burial habits of historic populations and individuals. Elemental spectra were performed from archeological samples of three chosen individuals from different time periods. The unusual presence of magnesium, aluminum, and silicon in the first sample could confirm the hypothesis of high degree of dental abrasion caused by particles from grinding stones in flour. In the second sample, presence of copper could confirm that bronze jewelery could lie near the buried body. The elemental composition of the third sample with the presence of lead and copper confirms the origin of individual to Napoleonic Wars because the damage to his teeth could be explained by the systematic utilization of the teeth for the opening of paper cartridges (a charge with a dose of gunpowder and a bullet), which were used during the 18th and the 19th century AD. All these results contribute to the reconstruction of life (first and third individual) and burial (second individual) habits of historic populations and individuals.

We characterized the morphological and anatomical adaptations of the lingual microstructures of the Eurasian collared dove and discussed their implications for its dietary niche. We analyzed tongues of nine S. decaocto using histological, histochemical, stereomicroscopic, and scanning electron microscopic techniques. Our findings showed that the tongue is relatively short with a tapered apex that carries a terminal lingual nail. However, the lingual body has median scales and is bordered laterally by filiform papillae. Further, the tongue body bears a distinctive papillary crest. The tongue root is nonpapillate and infiltered with orifices of the posterior salivary glands. The bulky laryngeal mound has a circular glottic fissure, carrying a single row of papillae at the rear edge. Concurrently, our histological and histochemical findings demonstrate that the tongue has taste buds, anterior and posterior salivary glands, along with an elongated entoglossum that extends from lingual apex to root. Besides, ovoid and globular mucous glands displayed intense alcianophilic reactions. More substantially, the palate is made up of three palatine ridges with a caudal choanal cleft that was bounded by two rows of palatine papillae. Our data indicate multiple and novel structural variations for the lingual and palatal sculptures coopted for their feeding style.

The carpenter ant Camponotus rufipes has intracellular bacteria in bacteriocytes scattered in the midgut epithelium, which have different amounts of endosymbionts, according to the developmental stages. However, there are no detailed data about the midgut cells in adult workers. The present work aimed to evaluate the morphology and cellular events that coordinate the abundance of endosymbionts in the midgut cells in C. rufipes workers. The midgut epithelium has digestive cells, bacteriocytes, and cells with intermediate morphology. The latter is similar to bacteriocytes, due to the abundance of endosymbionts, and similar to digestive cells, due to their microvilli. The digestive and intermediate cells are rich in autophagosomes and autolysosomes, both with bacteria debris in the lumen. These findings suggest that midgut cells of C. rufipes control the endosymbiont level by the autophagy pathway.