Microscopy Research and Technique最新文献

Segmentation and tracking are essential preliminary steps in the analysis of almost all live cell imaging applications. Although the number of open-source software systems that facilitate automated segmentation and tracking continue to evolve, many researchers continue to opt for manual alternatives for samples that are not easily auto-segmented, tracing cell boundaries by hand and reidentifying cells on consecutive frames by eye. Such methods are subject to inter-user variability, introducing idiosyncrasies into the results of downstream analysis that are a result of subjectivity and individual expertise. The methods are also susceptible to intra-user variability, meaning findings are challenging to reproduce. In this pilot study, we demonstrate and quantify the degree of intra- and inter-user variability in manual cell segmentation and tracking by comparing the phenotypic metrics extracted from cells segmented and tracked by different members of our research team. Furthermore, we compare the segmentation results for a ptychographic cell image obtained using different automated software and demonstrate the high dependence of performance on the imaging modality they were developed to handle. Our results show that choice of segmentation and tracking methods should be considered carefully in order to enhance the quality and reproducibility of results.

Analysis of indentation data of heterogeneous material, in particular, layer on an elastic substrate requires information about the contact area that is essential for calculating mechanical properties. The actual shape of the AFM-tip is not described by simple body of revolution. In this work, the indentation of a stiff layer on a hyperelastic substrate by a truncated conical tip is studied using finite element methods. The size of the tip, elastic modulus, and thickness of the layer are varied. The obtained model dependences of loading and contact area versus the depth of indentation are used in analysis of the experimental data of AFM indentation of stiff inhomogeneous nanolayers of polyurethane surface. Thickness and elastic modulus of the layers, fracture properties of the surface are studied. The obtained results can find application in the study of mechanical properties and fracture toughness of thin flexible films on an elastic substrate.

Currently, nanotechnology (NT) and nanoparticles (NPs) have gained significant attention in the scientific field due to their diverse application history. Particularly, in environmental applications, their antibacterial efficiency in fisheries due to antibacterial resistance. However, the NPs have been found toxic in the environment. Therefore, the current study aimed to fabricate less toxic NPs using environmentally dried leaves to check their antibacterial efficacy and possible toxicity against grass carp. The findings confirmed the good dispersity of obtained AgNPs, which further showed promising antibacterial activity against several bacterial isolates including Staphylococcus with a zone of inhibition (23.73 ± 0.57 nm). Also, the AgNPs were exposed to the grass carp (Ctenopharyngodon idella) for possible toxicity and toxic effects. First, the bioaccumulation of AgNPs was significantly observed in gills followed by intestines and muscles (p < 0.05). Finally, the AgNPs mainly accumulate in the liver, followed by the intestine, gills, and muscles. Additionally, the deposition of AgNPs in various organs resulted in histological alteration such as necrosis and infiltration of red blood cells in the intestine and the fusion of gill lamella. Hence, the synthesized NPs using dried leaf extract could be a promising approach in applied science. The significant features of the nanoparticles in the present work using green synthesis can help in synthesizing less toxic materials.

As a flexible biomolecule, the spatial structure of DNA is variable. The effects of concentration, metal cations, and low pH on DNA morphology were studied. For the high concentration of DNA, the cross-linked branch-like or network structures were formed. For the low concentration of DNA, isolated, random and freely loose linear DNA chains were presented. These phenomena were related to the intermolecular interactions. Branch-like DNA structures were reformed with the addition of metal cations to the low concentration of DNA at pH 7-4, suggesting the negative charges of DNA were neutralized, thus transforming the spatial structure of DNA into a low charge density morphology and presenting the hypochromic effect. Compared to the monovalent alkaline metal cations, more negative charges of DNA were screened by the alkaline-earth metal cations. Distinct DNA morphologies were observed for pH 3. The linear and condensed DNA structures were simultaneously observed, which was met regardless of the solution with or without the addition of metal cations. This was further confirmed by the absorbance of DNA. Compared to the pure DNA, bulky and aggregated DNA collapsed structures were formed when the sodium and magnesium cations were added to the reaction solution. In addition, it was verified that the condensed DNA structures failed to revert back to the chain structure by neutralizing acidic solutions with alkali, but the compacted DNA spheres became loose. The conductivities of various DNA morphologies were measured. They were morphology-dependent. This study provides guidance for the behavior of DNA in the acidic solutions and further promotes the application of DNA in DNA-based nano-optoelectronic devices.

Fluorescence imaging stands as a pivotal component in biomedical research, requiring the elimination of out-of-focus background noise resulting from wide-field volumetric illumination of the whole field-of-view and scattering within thick biological tissues. Traditional methods struggle to effectively address varying degrees of defocusing in fluorescence images. This study introduces the utilization of upU-Net, 3D U-Net, and 3D upU-Net as defocusing networks tailored for 2D and 3D wide-field fluorescence images, yielding notable enhancements. These advancements facilitate more economically viable confocal microscopy, delivering significant advantages to biologists presently utilizing wide-field fluorescence microscopy.

Ovarian cancer belongs to the category of gynecological malignancies and unfortunately holds the distinction of being the most aggressive among them. It is ranked as the fifth highest cause of cancer-related deaths in women worldwide. The utilization of metal nanoparticles (NPs) linked with natural herbal molecules in biomedical applications has been on the rise. Thymol carbon nanodot functionalized silver nanoparticles (ThCND-AgNPs) were synthesized in an original manner and subjected to thorough characterization, including analysis of their size, morphology, and elemental composition. The aim of this study is to investigate the effects of the ThCND-AgNPs on cell proliferation, invasion, and apoptotic gene expressions in OVCAR-3 ovarian cancer cells. The effect of ThCND-AgNPs on cell viability in OVCAR cells was determined in a dose- and time-dependent manner using the XTT method. The effect on the expression changes of apoptotic-related genes was assessed through the Real-time PCR method, while the anti-invasive activity was measured using the matrigel invasion chamber assay. The ThCND-AgNP molecule exhibited a dose- and time-dependent reduction in cell proliferation in OVCAR-3 cells. The IC50 values were determined to be 388.53 μg/mL at 24 h and 145.683 μg/mL at 48 h. Furthermore, the molecule was found to reduce cell invasion by 51.12% compared with the control group in OVCAR-3 cells. In terms of apoptotic-related genes, Bcl-2 expression was downregulated, while BAX, CASPASE-3, -8, and -9 expressions were unregulated. In conclusion, the obtained data reveal the potential antiproliferative, apoptotic, and anti-invasive effects of our original ThCND-AgNP molecule in ovarian cancer. While these results need further confirmation through more detailed experiments, they will provide insights for future studies.

The red-eared slider turtle, a species facing environmental challenges and habitat loss, exhibits a complex skin architecture that is crucial for its adaptation and survival. Our study aims to provide a comprehensive characterization of the turtle's skin structure and to elucidate the distribution and localization of its various cellular components, with a focus on understanding the skin's role in adaptation and ecological interactions. To achieve these goals, we employed light microscopy, transmission electron microscopy (TEM), and comprehensive immunofluorescence using 10 specific antibodies. The forelimb skin displays large- and moderate-sized scales with variations in color, including dark, yellow, and gray hues, likely contributing to camouflage and protection. The skin consists of corneous material, the epidermis, the dermis, and the hypodermis. The stratum basalis, stratum spinosum, and peri-corneous layer constitute the three distinct layers of the epidermis. There are four distinct types of chromatophores, including melanocytes located in the epidermis, while melanophores, xanthophores, and iridophores are found within the dermal layer. The skin also exhibits well-developed peripheral nerves, blood vessels, and subcutaneous muscles. Immunofluorescence staining further elucidates the distribution and localization of various skin cells. E-cadherin and CK14 are strongly expressed in the epidermal layers, excluding the corneous material. E-cadherin surrounds keratinocyte cells in the epidermis, facilitating cell-cell adhesion, while CK14 is present inside the keratinocyte cells, contributing to their internal structural integrity. Sox10 and CD117 identify the four chromatophore types, with Melan-A specifically detecting only melanocytes and melanophores and not labeling xanthophores and iridophores. Tom20 is used to detect mitochondrial distribution and intensity in the skin, revealing a high density of mitochondria in all epidermal layers, especially in melanocytes and melanophores, compared to xanthophores and iridophores. Numerous telocytes, spindle-shaped with extensions called telopods, are detected in the dermis using CD34, PDGFRα, and vimentin. The skin of the red-eared slider also shows abundant myofibroblasts and well-developed vascularization, with numerous blood vessels detected using α-SMA. This novel study offers an in-depth examination of the limb skin of the red-eared slider through the use of 10 distinct antibodies, uncovering the intricate interactions among its cellular components and providing valuable insights into its anatomical structure and physiological adaptations. Our findings contribute to a better understanding of the turtle's skin, which may aid in its conservation and management.

The aim of this work was to develop an ultrasonic-assisted synthesis method for the fabrication of CeO₂-doped Zr nanoparticles that would improve the performance of supercapacitor electrodes. This method, which eliminates the need for high-temperature calcination, involves embedding CeO₂ into Zr nanoparticles through 1 hr (CeO₂-Zr-1) and 2 hrs (CeO₂-Zr-2) of ultrasonic irradiation, resulting in the formation of nanostructures with significant improvements in their electrochemical properties. Through physicochemical analysis, we observed that the CeO₂-doped Zr nanoparticles, particularly those treated for 2 hrs (CeO₂-Zr-2), exhibit superior crystalline phase purity, optimal chemical surface composition, minimal agglomeration with particle sizes below 50 nm, and an impressive average surface area of 178 m²/g. Compared to the 1 hr irradiation samples (CeO₂-Zr-1) and undoped CeO₂ nanoparticles, the (CeO₂-Zr-2) electrodes demonstrated a remarkable capacitance of 198 Fg^-1 at a current density of 1 A/g while maintaining ~94.9% of their capacity after 3750 cycles. This indicates not only good reversibility but also exceptional stability. In (CeO₂-Zr-2) samples, the nanospherical structure achieved through ultrasonic synthesis is responsible for the enhanced capacitive behavior and stability, along with the synergistic effects caused by Zr doping, which improves the CeO₂ nanoparticle conductivity to a significant extent. Surface areas of the electrodes are larger due to the combination of these two materials, which contribute to their superior performance.

Cucumis anguria L. and Cucumis dipsaceus Ehrenb. ex Spach belong to the Cucurbitaceae family and are popularly known as "maxixe." In folk medicine, they are used to treat pneumonia, hyperglycemia, wounds, and malaria. This study aims to characterize the anatomy, histochemistry, and phytochemistry of C. anguria and C. dipsaceus. Conventional methods in plant anatomy and microscopy were used to prepare and analyze semi-permanent slides containing cross-sections of the stem, petiole, leaves, and paradermal sections of the leaf blade. For the histochemical analysis, different reagents were used according to the target metabolite. Phytochemical tests of methanolic extracts of leaves were performed by thin-layer chromatography. The microscopic analysis allowed obtaining the differentiation of plant cells, the presence of tector and glandular trichomes, in addition to showing anatomical traits such as stem shape, petiole shape, and organization of the mesophyll. Histochemistry showed the presence of alkaloids, starch, phenolic compounds, lipophilic compounds, lignin, and tannins. Phytochemical prospection identified monoterpenes, sesquiterpenes, triterpenes, steroids, alkaloids, and reducing sugars. The obtained results provide important information for quality control and species differentiation since there are few studies in the literature on these species.