Microscopy Research and Technique最新文献

Reactive lymphocytes are an important type of leukocytes, which are morphologically transformed from lymphocytes. The increase in these cells is usually a sign of certain virus infections, so their detection plays an important role in the fight against diseases. Manual detection of reactive lymphocytes is undoubtedly time-consuming and labor-intensive, requiring a high level of professional knowledge. Therefore, it is highly necessary to conduct research into computer-assisted diagnosis. With the development of deep learning technology in the field of computer vision, more and more models are being applied in the field of medical imaging. We aim to propose an advanced multi-object detection network and apply it to practical medical scenarios of reactive lymphocyte detection and other leukocyte detection. First, we introduce a space-to-depth convolution (SPD-Conv), which enhances the model's ability to detect dense small objects. Next, we design a dynamic large kernel attention (DLKA) mechanism, enabling the model to better model the context of various cells in clinical scenarios. Lastly, we introduce a brand-new feature fusion network, the asymptotic feature pyramid network (AFPN), which strengthens the model's ability to fuse multi-scale features. Our model ultimately achieves mAP50 of 0.918 for reactive lymphocyte detection and 0.907 for all leukocytes, while also demonstrating good interpretability. In addition, we propose a new peripheral blood cell dataset, providing data support for subsequent related work. In summary, our work takes a significant step forward in the detection of reactive lymphocytes.

Green synthesis of nanoparticles (NPs) is preferred for its affordability and environmentally friendly approach. This study explored the synthesis and characterization of silver NPs (AgNPs) and examined their impact on the growth of Zea mays, both alone and in combination with nickel chloride (NiCl₂). A methanolic leaf extract was combined with silver nitrate to synthesize AgNPs. Characterization of NPs was carried out through UV-vis spectroscopy, FT-IR, x-ray diffraction (XRD), energy-dispersive spectroscopy (EDS), and scanning electron microscopy (SEM). Eleven treatments (T1-T11) were made, and Z. mays seeds were subjected to NiCl₂ in pots after being soaked in AgNPs solution. Treatments were arranged to evaluate the effects of NiCl₂ (T1-T3), AgNPs (T4 and T5), and the interactive effects of AgNPs and NiCl₂ (T6-T11) on the planted seeds. UV-vis peaks at 410 nm confirmed the presence of AgNPs. The crystalline nature of AgNPs was confirmed through XRD analysis, and the presence of functional groups from biomolecules and capping agents was shown in FT-IR. The morphology of the NPs and elemental analysis were conducted using SEM and EDS, respectively. The size of the NPs was found 25-50 nm using Nano Measurer software. Growth inhibition was noticed in NiCl₂-treatments T1-T3. Maximum growth and 100% seed germination were observed in NP-treated seeds (T4 and T5). These two treatments also showed the highest germination index, root/shoot growth, and fresh/dry weights. In treatments T6-T11, the interaction between NiCl₂ and AgNP-soaked seeds showed that while AgNP concentrations alone promoted growth, this enhancement was suppressed by the presence of NiCl₂ in the soil. The inhibited values of T6-T11 were still greater than the control, indicating that soaking Z. mays seeds in AgNPs enhanced growth and mitigated nickel stress in the soil. Pigments, carbohydrates, and protein contents were highest in T4 and T5, whereas NiCl₂ reduced these values. Synthesized AgNPs could enhance Z. mays growth and reduce nickel stress at the applied amounts. Further investigation is required to determine the mechanisms of action of AgNPs and NiCl₂ in enhancing or reducing Z. mays seedling growth and yield.

The laryngeal mound (LM) formed the caudal part of the pharyngeal floor, which varied in position, shape, and length at different ages. This work aimed to study the morphogenesis of the LM in the embryonic and post hatching periods grossly, histologically, and by scanning electron microscopy using forty-eight Japanese quails. The LM primordia appeared on the 8th day of incubation as a raised elevation carried on a deep median symmetrical sulcus (glottis primordium). As a result of rapid differential LM parts growth, LM took different shapes with advanced ages, finally ending in a heart shape. Internally, LM was supported by hyaline laryngeal cartilages; a C-shaped cricoid cartilage that had two wings, paired fork-like two arytenoids, and a comma-shaped procricoid that had four articulations. The glottis appeared as a central longitudinal opening on the 13th day of incubation. With age advancing, it was characterized as a wide rostral commissure and a caudal narrow one that was supported on either side by arytenoid cartilages. Additionally, on the 13th day, a bilateral sagittal row to the glottis developed, consisting of 8-9 small caudally directed papillae. At that time, rostral and caudal transverse laryngeal papillary rows appeared. LM had compound tubuloalveolar submucosal laryngeal glands that were situated between M. dilator glottidis and cricohyoideus and opened on the dorsal surface of LM. Histochemically, the early post-hatching stages of the glandular secretion were PAS-positive while late post-hatching ages had alcinophilic reactions. In conclusion, the LM had rapid morphological developmental events in the early ages other than the adult ages.

Microscopic imaging aids disease diagnosis by describing quantitative cell morphology and tissue size. However, the high spatial resolution of these images poses significant challenges for manual quantitative evaluation. This project proposes using computer-aided analysis methods to address these challenges, enabling rapid and precise clinical diagnosis, course analysis, and prognostic prediction. This research introduces advanced deep learning frameworks such as squeeze-and-excitation and dilated dense convolution blocks to tackle the complexities of quantifying small and intricate breast cancer tissues and meeting the real-time requirements of pathological image analysis. Our proposed framework integrates a dense convolutional network (DenseNet) with an attention mechanism, enhancing the capability for rapid and accurate clinical assessments. These multi-classification models facilitate the precise prediction and segmentation of breast lesions in microscopic images by leveraging lightweight multi-scale feature extraction, dynamic region attention, sub-region classification, and regional regularization loss functions. This research will employ transfer learning paradigms and data enhancement methods to enhance the models' learning further and prevent overfitting. We propose the fine-tuning employing pre-trained architectures such as VGGNet-19, ResNet152V2, EfficientNetV2-B1, and DenseNet-121, modifying the final pooling layer in each model's last block with an SPP layer and associated BN layer. The study uses labeled and unlabeled data for tissue microscopic image analysis, enhancing models' robust features and classification abilities. This method reduces the costs and time associated with traditional methods, alleviating the burden of data labeling in computational pathology. The goal is to provide a sophisticated, efficient quantitative pathological image analysis solution, improving clinical outcomes and advancing the computational field. The model, trained, validated, and tested on a microscope breast image dataset, achieved recognition accuracy of 99.6% for benign and malignant secondary classification and 99.4% for eight breast subtypes classification. Our proposed approach demonstrates substantial improvement compared to existing methods, which generally report lower accuracies for breast subtype classification ranging between 85% and 94%. This high level of accuracy underscores the potential of our approach to provide reliable diagnostic support, enhancing precision in clinical decision-making.

When protein molecules come into contact with different types of substrate materials, the surface properties of the substrate will have a significant effect on their self-assembly behavior. The purpose of this study was to investigate the self-assembly behavior of zein molecules on the two different substrates. Herein, the microstructure of zein molecules on the surface of two typical substrates, mica and glass, were characterized in detail by atomic force microscopy. It was found that zein molecules self-assemble to form spherical structures with uniform size and close arrangement on mica substrates. Compared with mica, the rough glass surface possesses a larger water contact angle, which leads to weaker interaction between zein molecules and its surface, thus enhancing the interaction between zein molecules. Therefore, the zein molecules on the glass substrate exhibit a distinct hierarchical arrangement of one large globule surrounded by many smaller ones. This work provides valuable information for further study of the self-assembly behavior of zein molecules on the substrate surface.

Waveguide evanescent field fluorescence microscopy (WEFF) is an evanescent-based microscopy that utilizes a confined thin film of light, around 100 nm, to image the plasma membrane of cells attached to a waveguide. Low photobleaching and low background besides its high axial resolution allows time-lapse imaging to investigate changes in cell morphology in the presence or absence of chemical agents. Both large field of view (FOV) and uniform illumination are very important while imaging cell-substrate contacts with an evanescent field. In the current study, we demonstrate that the WEFF microscope is capable of large FOVs with a uniform illumination source and imaging over a very long time period with a simple and inexpensive experimental setup. The interaction of the trypsin with plasma membranes of live osteoblast cells is investigated. To analyze cell images (250 images), instead of relying on manual tracking, which is time-consuming and can introduce numerous errors, we performed image processing using TrackMate to investigate the dynamic response of cells upon exposure to trypsin. This helps to save time and increase the accuracy of the analysis. The powerful tracking and analysis capabilities of the TrackMate plugin in ImageJ are used to automatically detect the cells border and trace each cluster of cells. The reduction in cell area is accompanied by a notable increase in mean intensity, reflecting changes in the intracellular environment. However, the background did not change during the experiment, which proves that the fluorescence material remains attached to the cell membrane and does not leak into the cell medium.

Cordia diffusa K.C. Jacob, known as Sirunaruvili, belonging to the family Boraginaceae, is a rare endemic species. The study aimed to document the morpho-anatomical and histochemical characteristics of C. diffusa to facilitate its identification, as it is a highly threatened steno-endemic plant. The plant is an evergreen, woody shrub with brownish-gray, lenticellate stems and oblong-oblanceolate with pubescent leaves. Flowers are yellowish-white, tubular, fragrant, and arranged in umbellate cyme. Fruits and drupe, turning from green to orange when ripe, with viscid edible pulp. Flowering and fruiting occur year-round. The anatomical results showed that the adaxial surface of leaves had more stomata than the abaxial leaf surface. The pubescent plant surface was due to unicellular trichomes in the leaf's epidermal layer, and some of the epidermal cells had cystoliths. Petioles and stems had ridges, unicellular trichomes, and collateral vascular bundles with prominent water storage cells. Root exhibited radial vascular arrangements, cortical region, endodermis, and exarch xylem, with secondary growth forming periderm layers. Sclerenchymatous fibers and lignin accumulation were observed in the stem and roots. The histochemical study showed the presence of carbohydrates, proteins, lipids, acidic and neutral lipids, phenolic compounds, lignified cell walls, and alkaloids in various regions of the plant cell. The micro-morphological properties of the plant exhibited several unique aspects for accurate taxonomic identification and species delimitation, and this work offers many valuable details to plant taxonomists for future research.

This study delves into the intriguing world of extrachromosomal DNA (ecDNA) in breast cancer, uncovering its pivotal role in cancer's aggressiveness and genetic variability. ecDNA, a form of circular DNA found outside chromosomes, is known to play a significant role in cancer progression by increasing oncogene expression. Focusing on two contrasting cell lines, MDA-MB-231 (triple-negative) and MCF-7 (Luminal-A), we utilized advanced microscopy and fluorescence techniques to detect and characterize ecDNA. Our findings reveal a stark difference: MDA-MB-231 cells, known for their high metastatic potential, exhibit a striking abundance of ecDNA, manifested as double minutes and single form with intense fluorescence signals. In contrast, the less aggressive MCF-7 cells harbor significantly fewer ecDNA. This disparity highlights the potential of ecDNA as a key player in cancer progression and a promising target for novel therapies. This research sheds light on the unseen genetic forces driving breast cancer and opens the door to new strategies in cancer treatment. Further research is necessary to understand the mechanisms of ecDNA formation and its role in different breast cancer subtypes.

Diatoms (Bacillariophyceae) are diverse and abundant microalgae, and many of them are found in marine intertidal environments. Here, we present a new marine epizoic diatom isolated from Euspira gilva Philippi in intertidal area of Beihai city, Guangxi Zhuang Autonomous Region, named Ceratanaulus alaris L.J. Huang, Y.H. Gao & C.P. Chen sp. nov. Using light microscope (LM) and scanning electron microscope (SEM), we observed and described the morphology of C. alaris in detail. The valve center of C. alaris features a rimoportula with an external tube. The most notable characters in C. alaris are wing-like structures on both sides of the central tube. These structures extend from the central tube to the horn-like protrusions at the valve apices, with fissures evenly distributed on the wing-like structures. In this study, we compared the morphological structures of the new species with other species in the genus Ceratanaulus Górecka, Witkowski, Dabek & Ashworth, enhancing the understanding of the ultrastructure of C. alaris and facilitating the differentiation of this species from closely related taxa. The genus, Ceratanaulus, is newly recorded in China, thereby expanding the known geographical distribution of the genus.

This study aimed to describe the morphological features and microstructure of the upper, lower, and third eyelids of the black-winged kite, Elanus caeruleus, and to characterize the organized lymphoid follicles and lymphocytes in the eyelid mucosa. Additionally, it aimed to illustrate the importance of the eye adnexa in the eye's immune protection. The upper, lower, and third eyelids display varying morphological differences that seem to be closely linked to the birds' way of life, indicating adjustments to their environment and eating behaviors. The black-winged kite has large, forward-facing eyes positioned under a bony shelf (lacrimal process) that shades them. Both eyelids have thick, pigmented edges and bear two rows of long, finely modified filoplume feathers, which increase at the anterior canthus. Melanocytes appear in the stratum basale of upper and lower eyelids, and Langerhans cells were observed. Aggregations of lymphatic cells were present under the conjunctival epithelium within the stroma of the lower eyelid (in the orbital zone near the tarsal plate), while they were absent in the upper eyelid and nictitating membrane. The density of goblet cells on the conjunctival surface of the upper eyelid and the nictitating membrane is higher than that of the lower eyelid. The structure of the skin and conjunctiva in both the upper and lower eyelids contains numerous defensive immune cells that maintain ocular safety and hydration. Additionally, the lower eyelid's CALT constitutes most of the CALT tissue in E. caeruleus and is recognized as part of the mucosal immune system.