Applied Microscopy最新文献

The cosmetic-tattoo industry is evolving every year and the microstructures of the equipment have the great potential for semi-permanent makeup applications. Present paper explores the materials and microparticles of semi-permanent makeup tattoo needles. The surface of the five-round-shader tattoo needle used in semi-permanent makeup process was examined by scanning electron microscopy, and its elemental composition was analyzed by energy-dispersive X-ray spectroscopy. The comparison of five-round-shader needles have undergone thorough observation: original five-round-shader needle and distorted five-round-shader needle. The diameter of the sharp and rounded needle tip was measured at approximately 6.80 μm, while the deformed needle tip was approximately 16 μm thick, about 2.5 times thicker than the rounded needle tip. Many rosette-shaped lead (Pb) particles and irregular clusters adhere to the welded areas and closely adjacent needle shaft surfaces. The lead particles have a diameter ranging from 4 μm to 5 μm and exhibit a grid-like structure with a consistent thickness of plate-like shape. The distorted structure of Pb in rosette-shaped formations is shown to have originated from the grinding and polishing processes during needle manufacturing. To produce sterilized tattoo needles, high-quality tattoo needle inspection processes are necessary to remove any unhygienic substances adhering to the needle surface.

Glass vials are the most widely used primary containers for the packaging of parenteral products due to their optical clarity, general inertness, and hermetic properties, but under certain circumstances, they can pose safety concerns. Most of these issues are related to the potential formation of glass particulates through delamination or precipitation, resulting from the chemical interaction between the drug product and the inner surface of the glass vial. Hence, it is imperative for pharmaceutical companies to conduct product-vial compatibility studies to determine the appropriate packaging/container closure system. To support this development activity, scientists need to develop analytical methods to detect subvisible glass particulates in parenteral products, along with the appropriate positive controls, to facilitate detection and identification. This paper outlines the utilization of coaxial/episcopic and oblique illumination microscopy, combined with spectroscopic techniques, to detect thin glass particulates generated from a modified procedure. It also showcases the importance of angle-dependent lighting in visualizing positive control samples containing thin glass particulates. The analytical microscopy techniques discussed in this paper can assist scientists in selecting suitable container closure systems for developing parenteral products.

Oxide-supported metal catalysts are essential components in industrial processes for catalytic conversion. However, the performance of these catalysts is often compromised in high temperature reaction environments due to sintering effects. Currently, a number of studies are underway with the objective of improving the metal support interaction (MSI) effect in order to enhance sintering resistance by surface modification of the oxide support, including the formation of inhomogeneous defects on the oxide support, the addition of a rare earth element, the use of different facets, encapsulation, and other techniques. The recent developments in in situ gas phase transmission electron microscopy (TEM) have enabled direct observation of the sintering process of NPs in real time. This capability further allows to verify the efficacy of the methods used to tailor the support surface and contributes effectively to improving sintering resistance. Here, we review a few selected studies on how in situ gas phase TEM has been used to prevent the sintering of catalyst NPs on oxide supports.

The sliding filament theory and the cross-bridge model have been fundamental in understanding muscle contraction. While the cross-bridge model explains the interaction between a single myosin head and actin filament, the native myosin molecule consists of two heads. This review explores the possibility and mechanism of two-headed binding in myosin II to the actin. Recent studies using electron tomography and resonance energy transfer have provided evidence in support of the occurrence of two-headed binding. The flexibility of the regulatory light chain (RLC) appears to play a significant role in enabling this binding mode. However, high-resolution structures of the RLCs in the two-headed bound state have not yet been reported. Resolving these structures, possibly through sub-tomogram averaging or single-particle analysis, would provide definitive proof of the conformational flexibility of RLCs and their role in facilitating two-headed binding. Further investigations are also required to address questions such as the predominance of two-headed versus single-headed binding and the influence of the state of each of the heads on the other. An understanding of the mechanism of two-headed binding is crucial for developing a comprehensive model of the cross-bridge cycle of the native myosin molecule.

Plant cells are uniquely characterized by exhibiting cell walls, pigments, and phenolic compounds, which can impede microscopic observations by absorbing and scattering light. The concept of clearing was first proposed in the late nineteenth century to address this issue, aiming to render plant specimens transparent using chloral hydrate. Clearing techniques involve chemical procedures that render biological specimens transparent, enabling deep imaging without physical sectioning. Drawing inspiration from clearing techniques for animal specimens, various protocols have been adapted for plant research. These procedures include (i) hydrophobic methods (e.g., Visikol™), (ii) hydrophilic methods (ScaleP and ClearSee), and (iii) hydrogel-based methods (PEA-CLARITY). Initially, clearing techniques for plants were mainly utilized for deep imaging of seeds and leaves of herbaceous plants such as Arabidopsis thaliana and rice. Utilizing cell wall-specific fluorescent dyes for plants and fungi, researchers have documented the post-penetration behavior of plant pathogenic fungi within hosts. State-of-the-art plant clearing techniques, coupled with microbe-specific labeling and high-throughput imaging methods, offer the potential to advance the in planta characterization of plant microbiomes.

In this study, we investigate the effectiveness of noise reduction in electron holography, based on the wavelet hidden Markov model (WHMM), which allows the reasonable separation of weak signals from noise. Electron holography observations from a Nd₂Fe₁₄B thin foil showed that the noise reduction method suppressed artificial phase discontinuities generated by phase retrieval. From the peak signal-to-noise ratio, it was seen that the impact of denoising was significant for observations with a narrow spacing of interference fringes, which is a key parameter for the spatial resolution of electron holography. These results provide essential information for improving the precision of electron holography studies.

Due to the unique properties of the silk fibroin (SF) made from silkworm, SF-based hydrogels have recently received significant attention for various biomedical applications. However, research on the SF-based hydrogels isolated from spider silks has been rtricted due to the limited collection and preparation of naïve silk materials. Therefore, this study focused on the microstructural characteristics of hydrogel scaffolds derived from two types of woven silk glands: the major ampullate gland (MAG) and the tubuliform gland (TG), in the orb-web spider Trichonephila clavate. We compared these spider glands with those of the silk fibroin (SF) hydrogel scaffold extracted from the cocoon of the insect silkworm Bombyx mori. Our FESEM analysis revealed that the SF hydrogel has high porosity, translucency, and a loose upper structure, with attached SF fibers providing stability. The MAG hydrogel displayed even higher porosity, as well as elongated fibrous structures, and improved mechanical properties: while the TG hydrogel showed increased porosity, ridge-like or wall-like structures, and stable biocapacity formed by physical crosslinking. Due to their powerful and versatile microstructural characteristics, the MAG and TG hydrogels can become tailored substrates, very effective for tissue engineering and regenerative medicine applications.

The neuroprotective effects of umbelliferone (UMB) were visualized in three-dimensional (3D) images on vesicle density changes of organotypic hippocampal slice tissues (OHSCs) induced by scopolamine by high voltage electron microscopy. Observations revealed that the number of vesicles decreased in OHSCs induced by scopolamine, and UMB was found to inhibit scopolamine-induced reduction in vesicles, resulting in an increase in vesicle count. These 3D models provide valuable insight for understanding the increase of synapse vesicles in hippocampal tissues treated with UMB.

There has been an increase in the demand for purified protein as a result of recent developments in the structural biology of myosin 2. Although promising, current practices in myosin purification are usually time-consuming and cumbersome. The reported increased actin to myosin ratio in smooth muscles adds to the complexity of the purification process. Present study outlines a streamlined approach to isolate smooth muscle myosin 2 molecules from actomyosin suspension of chicken gizzard tissues. The procedure entails treating actomyosin for a brief period with actin-binding peptide phalloidin, followed by co-sedimentation and short column size exclusion chromatography. Typical myosin molecule with heavy and light chains and approximately 95% purity was examined using gel electrophoresis. Negative staining electron microscopy and image processing showed intact 10S myosin 2 molecules, proving that phalloidin is effective at eliminating majority of actin in the form of F-actin without dramatic alteration in the structure of myosin. The entire purification discussed here can be completed in a few hours, and further analysis can be done the same day. Thus, by offering quick and fresh supplies of native myosin molecules suited for structural research, specially cryo-electron microscopy, this innovative approach can be adapted to get around the drawbacks of time-intensive myosin purifying processes.