Microscopy Research and Technique最新文献

Bioglasses are solid materials consisted of sodium oxide, calcium oxide, silicon dioxide and phosphorus in various proportions and have used in bone tissue engineering. There have been ongoing efforts to improve the surface properties of bioglasses to increase biocompatibility and performance. The aim of the present study is to modify the bioglass surface with an amino acid mixture consisting of arginine, aspartic acid, phenylalanine, cysteine, histidine and lysine, to characterize the surface, and to evaluate the performance and biocompatibility in vitro and in vivo. The untreated bioglass, bioglass kept in simulated body fluid (SBF), and modified bioglass were used in further evaluation. After confirmation of the surface modification with FT-IR analyses and SEM analyses, MC3T3-E1 preosteoblasts adhesion on the surface was also revealed by SEM. The modified bioglass had significantly higher ALP activity in colorimetric measurement, rate of calcium accumulations in Alizarin red s staining, lower rate of cell death in Annexin-V/PI staining to determine apoptosis and necrosis. Having higher cell viability rate in MTT test and absence of genotoxicity in micronucleus test (OECD 487), the modified bioglass was further confirmed for biocompatibility in vitro. The results of the rat tibial defect model revealed that the all bioglass treatments had a significantly better bone healing score compared to the untreated negative control. However, the modified bioglass exhibited significantly better bone healing efforts especially during the first and the second months compared to the other bioglass treatment treatments. As a result, the amino acid surface modification of bioglasses improves the surface biocompatibility and osteogenic performance that makes the amino acid modified bioglass a better candidate for bone tissue engineering. RESEARCH HIGHLIGHTS: Bioglass surface modification with amino acids contributes to bioglass-tissue interaction with an improved cell attachment. Modified bioglass increases in vitro Alp activity and calcium accumulation, and also positively affects cell behavior by supporting cell adaptation. Bioglass exerts osteogenic potential in vivo especially during early bone healing.

Electron microscopy is essential for examining materials and biological samples at microscopic levels, providing detailed insights. Achieving high-quality imaging is often challenged by the potential damage high-energy beams can cause to sensitive samples. This study compares scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to evaluate image quality, noise levels, and the ability to preserve delicate specimens. We used a modified SEM system with a transmitted electrons conversion accessory, allowing it to operate like a TEM but at lower voltages, thereby reducing sample damage. Our analysis included quantitative assessments of noise levels and texture characteristics such as entropy, contrast, dissimilarity, homogeneity, energy, and correlation. This comprehensive evaluation directly compared traditional TEM and the adapted SEM system across various images. The results showed that TEM provided images with higher clarity and significantly lower noise levels, reinforcing its status as the preferred method for detailed studies. However, the modified SEM system also produced high-quality images at very low acceleration voltages, which is crucial for imaging samples sensitive to high-energy exposure. The texture metrics analysis highlighted the strengths and limitations of each method, with TEM images exhibiting lower entropy and higher homogeneity, indicating smoother and more uniform textures. This study emphasizes the importance of selecting the appropriate electron microscopy method based on research needs, such as sample sensitivity and required detail level. With its conversion accessory, the modified SEM system is a versatile and valuable tool, offering a practical alternative to TEM for various applications. This research enhances our understanding of the capabilities and limitations of SEM and TEM. It paves the way for further innovations in electron microscopy techniques, improving their applicability for studying sensitive materials. RESEARCH HIGHLIGHTS: Our study introduces a modified SEM adapter enabling TEM-like imaging at reduced voltages, effectively minimizing sample damage without compromising image resolution. Through comparative analysis, we found that images from the modified SEM closely match the quality of traditional TEM, showcasing significantly lower noise levels. This advancement underscores the SEM's enhanced capability for detailed structural analysis of sensitive materials, broadening its utility across materials science and biology.

Diabetes mellitus causes impaired diabetic wounds which is linked to a number of pathological alterations that impede the healing of wounds. In the current research, Swiss albino mice were given alloxan monohydrate to induce diabetes and excision wounds of approximately 6 mm using biopsy punch. The diabetic wounds were treated with various biomaterials including Vachellia nilotica extract (VN), Nigella sativa extract (NS), V. nilotica nanoparticles (VNNPs) and N. sativa nanoparticles (NSNPs). Their effects were determined by evaluating the percent wound contraction, healing time, and histopathological analysis. The serum level of various biochemical parameters that is, pro-inflammatory cytokines,  Matrix metalloproteinases (MMPs) and tissue inhibitor matrix metalloproteinases (TIMPs) were also determined. VNNPs group provided the best outcomes, with wound contraction 100% on 12^th day. According to histopathological examination, VNNPs group reduced inflammation and encouraged the formation of blood vessels, fibroblasts, and keratinocytes. VNNPs group significantly alleviated the serum level of pro-inflammatory cytokines that are, TNF-α (19.4 ± 1.5 pg/mL), IL-6 (13.8 ± 0.6 pg/mL), and IL-8 (24.8 ± 1.2 pg/mL) as compared with the diabetic mice. The serum level of MMP2 (248.2 ± 7.9 pg/mL), MMP7 (316 ± 5.2 pg/mL), and MMP9 (167.8 ± 12.1 pg/mL) in the same group VNNPs were also observed much less than the diabetic mice. The serum level of TIMPs (176.8 ± 2.9 pg/mL) in the VNNPs group was increased maximally with respect to diabetic mice. It is concluded that nanoparticles and biomaterials possess healing properties and have the ability to repair the chronic/diabetic wound. RESEARCH HIGHLIGHTS: UV-spectrophotometric and Fourier transform infrared spectroscopy observation for functional group analysis and possible linkage between conjugates Optimization of the histopathological and biochemical markers after application of the formulations Microscopic analysis of epithelial tissues for evaluation of healing mechanisms Speedy contraction of wounds as the alleviation of the inflammatory and necrotic factors.

Ultrasound images are susceptible to various forms of quality degradation that negatively impact diagnosis. Common degradations include speckle noise, Gaussian noise, salt and pepper noise, and blurring. This research proposes an accurate ultrasound image denoising strategy based on firstly detecting the noise type, then, suitable denoising methods can be applied for each corruption. The technique depends on convolutional neural networks to categorize the type of noise affecting an input ultrasound image. Pre-trained convolutional neural network models including GoogleNet, VGG-19, AlexNet and AlexNet-support vector machine (SVM) are developed and trained to perform this classification. A dataset of 782 numerically generated ultrasound images across different diseases and noise types is utilized for model training and evaluation. Results show AlexNet-SVM achieves the highest accuracy of 99.2% in classifying noise types. The results indicate that, the present technique is considered one of the top-performing models is then applied to real ultrasound images with different noise corruptions to demonstrate efficacy of the proposed detect-then-denoise system. RESEARCH HIGHLIGHTS: Proposes an accurate ultrasound image denoising strategy based on detecting noise type first. Uses pre-trained convolutional neural networks to categorize noise type in input images. Evaluates GoogleNet, VGG-19, AlexNet, and AlexNet-support vector machine (SVM) models on a dataset of 782 synthetic ultrasound images. AlexNet-SVM achieves highest accuracy of 99.2% in classifying noise types. Demonstrates efficacy of the proposed detect-then-denoise system on real ultrasound images.

The pollen morphology of 33 species belonging to the genera Lactuca, Launaea, Cicerbita, Sonchus, Condrilla (tribe Cichorieae: Asteraceae) from Iran were studied with light and electron microscopies. The pollen grains of the investigated taxa are almost oblate-spherical to prolate-spherical, the shapes are circular in equatorial view, hexagonal in polar view, only Sonchus species are triangular or quadrangular in polar view, isopolar, symmetrical and trizonocolporate, except Sonchus and Chondrilla species, which have tri- to tetrazonocolporate pollen grains, and echinolophate. The size and number of spines in the polar area and the number of spines in the paraporal ridges and equatorial ridges proved to be the most taxonomically authentic characteristics in this study. Three pollen types were distinguished, as in alignment with previous studies. Cluster analysis by Ward method based on 10 quantitative morphological characteristics of pollen grains revealed that the species could be grouped based on the pollen characteristics. Moreover, three species of Cicerbita including Cicerbita scoparia (Rech. f. & Köie) Kitam, C. rechingeriana (Tuisl) Coșkunç., M. Güzel & N. Kilian and C. microcephala (DC.) M. Güzel, Coșkunç. & N. Kilian were recorded for the first time in Iran. Although the characteristics of the pollen grains were useful for distinguishing the species in sub-tribes, in some cases, this did not confirm recent phylogenetic results. The pollen characteristics described in this study provide data that may be useful for future systematic studies aiming to complete the knowledge on Cichorieae species. RESEARCH HIGHLIGHTS: In the present study, the pollen morphology of 33 species belonging to five genera of the Cichorieae are investigated with scanning electron microscopy and 12 species are analyzed for the first time. The cluster analysis based on the quantitative morphological characteristics of pollen grains revealed that the species could be grouped based on the pollen characteristics. In addition, three species of Cicerbita are reported for the first time in Iran.

The most prominent insect pest harming eucalyptus trees in China was Endoclita vietnamensis, which resulted in considerable losses to the environment and economy. This study examined the hair brush on the postpedes of male E. vietnamensis using transmission and scanning electron microscopy. The finding revealed that the hair brush was made up of many large, flattened, pale yellow, specialized scales that were highly elongated. These scales were firmly affixed to the epidermis at their base, and at the attachment site, there was a swollen hair follicle. The interior of the pencil was hollow with ducts, and the surface featured rows of pores and longitudinal ridges. Numerous features, including mitochondria, lipid droplets, smooth and rough endoplasmic reticulum, and microvilli, were present in the tissue cells of the tibial segment of postpedes of male moths. The tibial segment of postpedes of male moths exhibited thicker epidermis, cavities, and voids in the tissue cortex, suggesting the presence of secretory activities in the tissue cells. With the location and structure of the pheromone gland determined, a theoretical foundation for future research into the communication between the female and male E. vietnamensis as well as the development of sex-specific pheromone-based catching and killing methods will be available. RESEARCH HIGHLIGHTS: Endoclita vietnamensis Buchsbaum & Grehan, 2022 is the most prominent insect pest harming eucalyptus trees in China. With the location and structure of the pheromone gland determined, a theoretical foundation for future research into the communication between the female and male E. vietnamensis as well as the development of sex-specific pheromone-based catching and killing methods will be available.

Sperm morphology is considered the best indicator of male fertility. In Neotropical bats, important aspects of sperm morphology have been scantly studied. The aim of the present study was to characterize and compare the sperm morphology and morphometry of Artibeus planirostris and Sturnira erythromos. A total of 11 specimens were analyzed from the Colección de Mamíferos Lillo: five A. planirostris and six S. erythromos. The fixed epididymis were extracted and macerated in Farmer's solution, followed by the routine cytological procedure with different stains. To carry out the description and morphometric analysis, microphotographs were taken under an optical, epifluorescence and scanning electron microscope. A total of 50 sperm from each individual were measured for morphometric analysis. The percentage of normal/abnormal spermatozoa was estimated and the sperm abnormalities were classified. Both species showed morphologically simple spermatozoa with a spatulate head, a short neck, a helical midpiece and a tail that tapers at the final end, similar to other species of Phyllostomidae. The differences observed were: apex of the head was conical in A. planirostris and was oval in S. erythromos; longer head and midpiece in S. erythomos and longer sperm in A. planirostris. Both species showed a high percentage of sperm with normal appearance: 65% for A. planirostris and 72% for S. erythromos. The main sperm abnormalities were: scattered tails and heads, coiled tails, folded midpieces and presence of cytoplasmic droplets. The present work will improve the understanding of their reproductive biology. RESEARCH HIGHLIGHTS: Morphological descriptions and morphometric analyses of the sperm of Artibeus planirostris and Sturnira erythromos were carried out with optical, epifluorescence and scanning electron microscopy.

Ultrafast fluorescent confocal microscopy is a hypothetical approach for breast cancer detection because of its potential to achieve instantaneous, high-resolution images of cellular-level tissue features. Traditional approaches such as mammography and biopsy are laborious, invasive, and inefficient; confocal microscopy offers many benefits over these approaches. However, confocal microscopy enables the exact differentiation of malignant cells, the expeditious examination of extensive tissue sections, and the optical sectioning of tissue samples into tiny slices. The primary goal should be to prevent cancer altogether, although detecting it early can help achieve that objective. This research presents a novel Breast Histopathology Convolutional Neural Network (BHCNN) for feature extraction and recursive feature elimination method for selecting the most significant features. The proposed approach utilizes full slide images to identify tissue in regions affected by invasive ductal carcinoma. In addition, a transfer learning approach is employed to enhance the performance and accuracy of the models in detecting breast cancer, while also reducing computation time by modifying the final layer of the proposed model. The results showed that the BHCNN model outperformed other models in terms of accuracy, achieving a testing accuracy of 98.42% and a training accuracy of 99.94%. The confusion matrix results show that the IDC positive (+) class achieved 97.44% accuracy and 2.56% inaccurate results, while the IDC negative (-) class achieved 98.73% accuracy and 1.27% inaccurate results. Furthermore, the model achieved less than 0.05 validation loss. RESEARCH HIGHLIGHTS: The objective is to develop an innovative framework using ultra-fast fluorescence confocal microscopy, particularly for the challenging problem of breast cancer diagnosis. This framework will extract essential features from microscopy and employ a gradient recurrent unit for detection. The proposed research offers significant potential in enhancing medical imaging through the provision of a reliable and resilient system for precise diagnosis of breast cancer, thereby propelling the progression of state-of-the-art medical technology. The most suitable feature was determined using BHRFE optimization techniques after retrieving the features by proposed model. Finally, the features chosen are integrated into a proposed methodology, which is then classified using a GRU deep model. The aforementioned research has significant potential to improve medical imaging by providing a complex and reliable system for precise evaluation of breast cancer, hence advancing the development of cutting-edge medical technology.

Fibraurea tinctoria Lour. (Menispermaceae) is a medicinal plant used by many local tribes in Indonesia and other countries. This species has pharmacological potential, such as antimalaria, antibacterial, antitumor, antioxidant, anticancer, antidiabetic, anti-inflammatory, and hepato-protector. However, the secretory structures and sites of secondary metabolite accumulation still need to be studied. This study aimed to explore the secretory structures and reveal the accumulation sites of the main classes of secondary metabolites in the root, stem, and leaves of F. tinctoria. Histochemical analysis using various specific reagents was performed on freehand sections of the fresh organs to detect alkaloids, phenolics, terpenoids, and lipophilic compounds. Observations of secretory structures were carried out using light microscopy. The result showed that idioblast was the predominant secretory structure in the plant organs and contained all the chemical groups investigated, while laticifer accumulated alkaloids were found only in the stem. Some secondary metabolites were localized in the xylem, intercellular space, and cuticle layer. Crystals were observed in the root, stem, and leaf of this species. These results might be helpful as guide information for extracting the particular plant parts to obtain the desired secondary metabolites. It suggested that leaves are a potential alternative source of medicinal raw material of this species, in addition to the stem. RESEARCH HIGHLIGHTS: The first study on secretory structures of Fibraurea tinctoria. Secondary metabolites in F. tinctoria organs accumulate in specialized and unspecialized structures. The idioblast is the primary secretory structure in the organs of F. tinctoria spreading over the root, stem, and leaves.

The present study unveils the intricate details on the morphology of thrips through optical, field emission scanning electron microscopy (FE-SEM) and mitochondrial cytochrome oxidase I gene-based molecular identification tools. The variation in the morphological characters namely, antennae (seven-segmented with forked sensorium on third, fourth segments), ctenidia (paired ctenidia were present in 5^th-8^th abdominal segments laterally), pronotum (two pairs of posteroangular setae) were observed in both Thrips tabaci and Thrips parvispinus, respectively. Similarly, ocelli color (brown and red colored), ocellar setae (two and three pairs of ocellar setae on the head of T. tabaci and T. parvispinus, respectively. Irregular reticulate striations on metascutum and medial striations are present in the metanotum of T. parvispinus; forewings with 6 distal setae in the first vein and 15 distal setae in the second vein in T. tabaci and forewings of T. parvispinus with complete rows of setae in the first and second vein in T. parvispinus; abdomen with median dorsal setae present in the tergite of T. tabaci and presence of 6-12 discal setae in sternites III-VI segments, absence of discal setae on sternites II and VII in T. parvispinus were observed, respectively. Further, FE-SEM studies revealed that similar type of sensilla namely, sensilla basiconica (SBI, SBII, SBIII), sensilla chaetica (SChI, SChII), sensilla trichodea (ST), sensilla campaniformia (SCa), and sensilla cavity (SCav) were recorded in both the species and variations were observed in length of above sensilla of T. tabaci and T. parvispinus. Additionally, Bohm bristles (Bb) and microtrichia (Mt) on the antennal surface contributed to a comprehensive understanding of their ultrastructural features. The molecular characterization revealed a single ~450 bp nucleotide fragment with over 98% similarity for the confirmation of T. tabaci and T. parvispinus in concurrence with NCBI data. RESEARCH HIGHLIGHTS: Microscopy-based morphological and ultrastructural characterization of Thrips tabaci Lindeman and Thrips parvispinus Karny.