Hana Duranova, Lenka Kuzelova, Petra Borotova, Veronika Simora, Veronika Fialkova
Human umbilical vein endothelial cells (HUVECs) are primary cells isolated from the vein of an umbilical cord, extensively used in cardiovascular studies and medical research. These cells, retaining the characteristics of endothelial cells in vivo, serve as a valuable cellular model system for understanding vascular biology, endothelial dysfunction, pathophysiology of diseases such as atherosclerosis, and responses to different drugs or treatments. Transmission electron microscopy (TEM) has been a cornerstone in revealing the detailed architecture of multiple cellular model systems including HUVECs, allowing researchers to visualize subcellular organelles, membrane structures, and cytoskeletal elements. Among them, the endoplasmic reticulum, Golgi apparatus, mitochondria, and nucleus can be meticulously examined to recognize alterations indicative of cellular responses to various stimuli. Importantly, Weibel-Palade bodies are characteristic secretory organelles found in HUVECs, which can be easily distinguished in the TEM. These distinctive structures also dynamically react to different factors through regulated exocytosis, resulting in complete or selective release of their contents. This detailed review summarizes the ultrastructural features of HUVECs and highlights the utility of TEM as a pivotal tool for analyzing HUVECs in diverse research frameworks, contributing valuable insights into the comprehension of HUVEC behavior and enriching our knowledge into the complexity of vascular biology.
人脐带静脉内皮细胞(HUVECs)是从脐带静脉中分离出来的原代细胞,广泛用于心血管研究和医学研究。这些细胞保留了体内内皮细胞的特征,是了解血管生物学、内皮功能障碍、动脉粥样硬化等疾病的病理生理学以及对不同药物或治疗的反应的重要细胞模型系统。透射电子显微镜(TEM)是揭示包括 HUVECs 在内的多种细胞模型系统详细结构的基石,它使研究人员能够直观地观察亚细胞器、膜结构和细胞骨架元素。其中,可以对内质网、高尔基体、线粒体和细胞核进行细致的检查,以识别细胞对各种刺激的反应变化。重要的是,Weibel-Palade 体是 HUVEC 中发现的特征性分泌细胞器,在 TEM 中很容易区分。这些独特的结构还能通过调节外渗对不同因素做出动态反应,从而完全或有选择性地释放其内容物。这篇详细的综述总结了 HUVEC 的超微结构特征,强调了 TEM 作为一种关键工具在不同研究框架下分析 HUVEC 的实用性,为理解 HUVEC 的行为提供了宝贵的见解,丰富了我们对复杂的血管生物学的认识。
{"title":"Human Umbilical Vein Endothelial Cells as a Versatile Cellular Model System in Diverse Experimental Paradigms: An Ultrastructural Perspective.","authors":"Hana Duranova, Lenka Kuzelova, Petra Borotova, Veronika Simora, Veronika Fialkova","doi":"10.1093/mam/ozae048","DOIUrl":"10.1093/mam/ozae048","url":null,"abstract":"<p><p>Human umbilical vein endothelial cells (HUVECs) are primary cells isolated from the vein of an umbilical cord, extensively used in cardiovascular studies and medical research. These cells, retaining the characteristics of endothelial cells in vivo, serve as a valuable cellular model system for understanding vascular biology, endothelial dysfunction, pathophysiology of diseases such as atherosclerosis, and responses to different drugs or treatments. Transmission electron microscopy (TEM) has been a cornerstone in revealing the detailed architecture of multiple cellular model systems including HUVECs, allowing researchers to visualize subcellular organelles, membrane structures, and cytoskeletal elements. Among them, the endoplasmic reticulum, Golgi apparatus, mitochondria, and nucleus can be meticulously examined to recognize alterations indicative of cellular responses to various stimuli. Importantly, Weibel-Palade bodies are characteristic secretory organelles found in HUVECs, which can be easily distinguished in the TEM. These distinctive structures also dynamically react to different factors through regulated exocytosis, resulting in complete or selective release of their contents. This detailed review summarizes the ultrastructural features of HUVECs and highlights the utility of TEM as a pivotal tool for analyzing HUVECs in diverse research frameworks, contributing valuable insights into the comprehension of HUVEC behavior and enriching our knowledge into the complexity of vascular biology.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141180070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lab managers and users of scanning electron microscope or electron probe microanalyzer facilities aiming for qualitative or quantitative X-ray analyses require comprehensive, yet flexible documentation structures for their daily work and available reference material, with a complete X-ray data library, a repository of energy- and wavelength-dispersive spectra, and an instrument scheduling mechanism. An online multilaboratory database system available at https://de-ma.ch is presented with the primary goals of providing information on microanalytical reference materials, analytical setups, characteristic X-ray data, and for managing reservation and training requests. This website is designed for multiuser facilities, where experience ranges from beginners to expert users. Registered users will find these tools useful in developing and maintaining high-quality, reproducible, and efficient analyses, whereas lab managers will keep records of their microanalytical reference material database and analytical protocols. The database also serves an educational purpose by (a) providing information on reference materials, (b) encouraging students to select appropriate X-ray lines to analyze, (c) providing analytical setups for point analysis or mapping, (d) identifying unknown X-ray lines, (e) displaying energy- or wavelength-dispersive spectra, and (f) recalculating mineral formula from quantitative wt% analysis results, based on a number of oxygen atoms or cations.
旨在进行定性或定量 X 射线分析的扫描电子显微镜或电子探针微分析设备的实验室管理人员和用户需要全面而灵活的文档结构来记录他们的日常工作和可用参考材料,包括完整的 X 射线数据图书馆、能量和波长色散光谱库以及仪器调度机制。https://de-ma.ch 上提供了一个在线多实验室数据库系统,其主要目标是提供有关微分析参考材料、分析设置、特征 X 射线数据的信息,以及管理预订和培训请求。该网站专为多用户设施而设计,用户经验从初学者到专家不等。注册用户会发现这些工具在开发和维护高质量、可重现和高效的分析方面非常有用,而实验室管理人员则会保存其微量分析参考材料数据库和分析协议的记录。该数据库还具有以下教育功能:(a) 提供有关参考材料的信息;(b) 鼓励学生选择适当的 X 射线进行分析;(c) 提供用于点分析或绘图的分析设置;(d) 识别未知的 X 射线;(e) 显示能量或波长色散光谱;(f) 根据氧原子或阳离子的数量,从重量百分比定量分析结果中重新计算矿物公式。
{"title":"De-MA.ch: A Web Database for Electron and X-ray Microanalysis to Assist Electron Microscope Lab Managers and Users.","authors":"Julien M Allaz","doi":"10.1093/mam/ozae051","DOIUrl":"10.1093/mam/ozae051","url":null,"abstract":"<p><p>Lab managers and users of scanning electron microscope or electron probe microanalyzer facilities aiming for qualitative or quantitative X-ray analyses require comprehensive, yet flexible documentation structures for their daily work and available reference material, with a complete X-ray data library, a repository of energy- and wavelength-dispersive spectra, and an instrument scheduling mechanism. An online multilaboratory database system available at https://de-ma.ch is presented with the primary goals of providing information on microanalytical reference materials, analytical setups, characteristic X-ray data, and for managing reservation and training requests. This website is designed for multiuser facilities, where experience ranges from beginners to expert users. Registered users will find these tools useful in developing and maintaining high-quality, reproducible, and efficient analyses, whereas lab managers will keep records of their microanalytical reference material database and analytical protocols. The database also serves an educational purpose by (a) providing information on reference materials, (b) encouraging students to select appropriate X-ray lines to analyze, (c) providing analytical setups for point analysis or mapping, (d) identifying unknown X-ray lines, (e) displaying energy- or wavelength-dispersive spectra, and (f) recalculating mineral formula from quantitative wt% analysis results, based on a number of oxygen atoms or cations.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141180098","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kingham [(1982). The post-ionization of field evaporated ions: A theoretical explanation of multiple charge states. Surf Sci116(2), 273-301] provided equations for the probability of observing higher charge states in atom probe tomography (APT) experiments. These "Kingham curves" have wide application in APT, but cannot be analytically transformed to provide the electric field in terms of the easily measured charge state ratio (CSR). Here we provide a numerical scheme for the calculation of Kingham curves and the variation in electric field with CSR. We find the variation in electric field with CSR is well described by a simple two- or three-parameter equation, and the model is accurate to most elements and charge states. The model is applied to experimental APT data of pure aluminium and a microalloyed steel, demonstrating that the methods described in this work can be easily applied to a variety of APT problems to understand electric field variations.
{"title":"Estimation of the Electric Field in Atom Probe Tomography Experiments Using Charge State Ratios.","authors":"Levi Tegg, Leigh T Stephenson, Julie M Cairney","doi":"10.1093/mam/ozae047","DOIUrl":"10.1093/mam/ozae047","url":null,"abstract":"<p><p>Kingham [(1982). The post-ionization of field evaporated ions: A theoretical explanation of multiple charge states. Surf Sci116(2), 273-301] provided equations for the probability of observing higher charge states in atom probe tomography (APT) experiments. These \"Kingham curves\" have wide application in APT, but cannot be analytically transformed to provide the electric field in terms of the easily measured charge state ratio (CSR). Here we provide a numerical scheme for the calculation of Kingham curves and the variation in electric field with CSR. We find the variation in electric field with CSR is well described by a simple two- or three-parameter equation, and the model is accurate to most elements and charge states. The model is applied to experimental APT data of pure aluminium and a microalloyed steel, demonstrating that the methods described in this work can be easily applied to a variety of APT problems to understand electric field variations.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141262389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alexandra L Day, Carolin B Wahl, Vishu Gupta, Roberto Dos Reis, Wei-Keng Liao, Chad A Mirkin, Vinayak P Dravid, Alok Choudhary, Ankit Agrawal
Traditionally, materials discovery has been driven more by evidence and intuition than by systematic design. However, the advent of "big data" and an exponential increase in computational power have reshaped the landscape. Today, we use simulations, artificial intelligence (AI), and machine learning (ML) to predict materials characteristics, which dramatically accelerates the discovery of novel materials. For instance, combinatorial megalibraries, where millions of distinct nanoparticles are created on a single chip, have spurred the need for automated characterization tools. This paper presents an ML model specifically developed to perform real-time binary classification of grayscale high-angle annular dark-field images of nanoparticles sourced from these megalibraries. Given the high costs associated with downstream processing errors, a primary requirement for our model was to minimize false positives while maintaining efficacy on unseen images. We elaborate on the computational challenges and our solutions, including managing memory constraints, optimizing training time, and utilizing Neural Architecture Search tools. The final model outperformed our expectations, achieving over 95% precision and a weighted F-score of more than 90% on our test data set. This paper discusses the development, challenges, and successful outcomes of this significant advancement in the application of AI and ML to materials discovery.
传统上,材料发现更多地依靠证据和直觉,而非系统设计。然而,"大数据 "的出现和计算能力的指数级增长重塑了这一格局。如今,我们利用模拟、人工智能(AI)和机器学习(ML)来预测材料特性,这大大加快了新型材料的发现。例如,在单个芯片上创建数百万个不同纳米粒子的组合巨库,激发了对自动表征工具的需求。本文介绍了一种 ML 模型,该模型专门用于对来自这些巨型库的纳米颗粒的灰度高角度环形暗场图像进行实时二元分类。鉴于与下游处理错误相关的高成本,我们模型的主要要求是在保持对未见图像的有效性的同时,最大限度地减少误报。我们详细阐述了计算方面的挑战和我们的解决方案,包括管理内存限制、优化训练时间和利用神经架构搜索工具。最终的模型超出了我们的预期,在测试数据集上达到了 95% 以上的精确度和 90% 以上的加权 F 分数。本文讨论了人工智能和 ML 在材料发现领域应用的这一重大进展的开发、挑战和成功结果。
{"title":"Machine Learning-Enabled Image Classification for Automated Electron Microscopy.","authors":"Alexandra L Day, Carolin B Wahl, Vishu Gupta, Roberto Dos Reis, Wei-Keng Liao, Chad A Mirkin, Vinayak P Dravid, Alok Choudhary, Ankit Agrawal","doi":"10.1093/mam/ozae042","DOIUrl":"10.1093/mam/ozae042","url":null,"abstract":"<p><p>Traditionally, materials discovery has been driven more by evidence and intuition than by systematic design. However, the advent of \"big data\" and an exponential increase in computational power have reshaped the landscape. Today, we use simulations, artificial intelligence (AI), and machine learning (ML) to predict materials characteristics, which dramatically accelerates the discovery of novel materials. For instance, combinatorial megalibraries, where millions of distinct nanoparticles are created on a single chip, have spurred the need for automated characterization tools. This paper presents an ML model specifically developed to perform real-time binary classification of grayscale high-angle annular dark-field images of nanoparticles sourced from these megalibraries. Given the high costs associated with downstream processing errors, a primary requirement for our model was to minimize false positives while maintaining efficacy on unseen images. We elaborate on the computational challenges and our solutions, including managing memory constraints, optimizing training time, and utilizing Neural Architecture Search tools. The final model outperformed our expectations, achieving over 95% precision and a weighted F-score of more than 90% on our test data set. This paper discusses the development, challenges, and successful outcomes of this significant advancement in the application of AI and ML to materials discovery.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140958450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shahar Seifer, Peter Kirchweger, Karlina Maria Edel, Michael Elbaum
4D STEM is an emerging approach to electron microscopy. While it was developed principally for high-resolution studies in materials science, the possibility to collect the entire transmitted flux makes it attractive for cryomicroscopy in application to life science and radiation-sensitive materials where dose efficiency is of utmost importance. We present a workflow to acquire tomographic tilt series of 4D STEM data sets using a segmented diode and an ultrafast pixelated detector, demonstrating the methods using a specimen of a T4 bacteriophage. Full integration with the SerialEM platform conveniently provides all the tools for grid navigation and automation of the data collection. Scripts are provided to convert the raw data to mrc format files and further to generate a variety of modes representing both scattering and phase contrasts, including incoherent and annular bright field, integrated center of mass, and parallax decomposition of a simulated integrated differential phase contrast. Principal component analysis of virtual annular detectors proves particularly useful, and axial contrast is improved by 3D deconvolution with an optimized point spread function. Contrast optimization enables visualization of irregular features such as DNA strands and thin filaments of the phage tails, which would be lost upon averaging or imposition of an inappropriate symmetry.
{"title":"Optimizing Contrast in Automated 4D STEM Cryotomography.","authors":"Shahar Seifer, Peter Kirchweger, Karlina Maria Edel, Michael Elbaum","doi":"10.1093/mam/ozae050","DOIUrl":"10.1093/mam/ozae050","url":null,"abstract":"<p><p>4D STEM is an emerging approach to electron microscopy. While it was developed principally for high-resolution studies in materials science, the possibility to collect the entire transmitted flux makes it attractive for cryomicroscopy in application to life science and radiation-sensitive materials where dose efficiency is of utmost importance. We present a workflow to acquire tomographic tilt series of 4D STEM data sets using a segmented diode and an ultrafast pixelated detector, demonstrating the methods using a specimen of a T4 bacteriophage. Full integration with the SerialEM platform conveniently provides all the tools for grid navigation and automation of the data collection. Scripts are provided to convert the raw data to mrc format files and further to generate a variety of modes representing both scattering and phase contrasts, including incoherent and annular bright field, integrated center of mass, and parallax decomposition of a simulated integrated differential phase contrast. Principal component analysis of virtual annular detectors proves particularly useful, and axial contrast is improved by 3D deconvolution with an optimized point spread function. Contrast optimization enables visualization of irregular features such as DNA strands and thin filaments of the phage tails, which would be lost upon averaging or imposition of an inappropriate symmetry.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141419824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Correction to: Automated Grain Boundary Detection for Bright-Field Transmission Electron Microscopy Images via U-Net.","authors":"","doi":"10.1093/micmic/ozad140","DOIUrl":"10.1093/micmic/ozad140","url":null,"abstract":"","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138830372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pollen micromorphological traits with taxonomic implications are first reported from the study area for 50 Asteraceous taxa belonging to nine tribes. Cichorieae (21 taxa), Cardueae (11 taxa), Inuleae (six taxa), and Anthemideae (four taxa) are the leading tribes. The research included Cousinia haeckeliae, Himalaiella afghana, Pterachaenia stewartii (endemic to Afghanistan and Pakistan), and Xylanthemum macropodum (endemic to Baluchistan). Light and scanning electron microscopy were employed for the visualization of pollen photomicrographs. The data was analyzed statistically via SPSS, PAST, and Origin. Significant diagnostic qualitative and quantitative palynological traits were explored for discrimination down to the species level within the tribes. All the investigated taxa possessed radial symmetry, isopolarity, and monad form (characters for distinction at the subdivision level). The aperture types were trizonocolporate, tetrazonocolporate, and tricolporate with number position and character (NPC) formulas N3P4C5, N4P4C5, and N3P4C3. Goniotreme, peritreme, and ptychotreme types of amb were recognized. Echinate, echinate lophate, scabrate, and gemmate sculpturing were present with and without perforated surface patterns. Variations in the shapes in polar and equatorial views and lacuna shapes further assisted the separation of taxa. The observed shape classes were perprolate, prolate spheroidal, prolate, subprolate, oblate spheroidal, suboblate, and oblate. Principal component analysis, correlation, standard probability plots, and ridge line paired features plot for quantitative variables determined the positive correlation between the length and width of colpi in equatorial and polar view with polar axis and equatorial diameter and number of spines between colpi with the number of spines per pollen. The number of spines per pollen was negatively correlated with the width and length of colpi in the polar view. Multiple sample analysis of variance (ANOVA) concluded that a high statistically significant difference exists among the means of analyzed traits. The examined qualitative and quantitative palynological traits revealed noticeable variations, thus providing the source for species discrimination in Asteraceous tribes.
{"title":"Systematic Implications of Palynomorph Diversity Using Microscopic Trends Among Asteraceous Flora From the Drylands of Baluchistan, Pakistan.","authors":"Bibi Sadia, Mushtaq Ahmad, Mohamed Fawzy Ramadan, Muhammad Zafar, Shazia Sultana, Wajia Noor, Trobjon Makhkamov, Akramjon Yuldashev, Khislat Khaydarov, Andrea Pieroni","doi":"10.1093/mam/ozae039","DOIUrl":"10.1093/mam/ozae039","url":null,"abstract":"<p><p>Pollen micromorphological traits with taxonomic implications are first reported from the study area for 50 Asteraceous taxa belonging to nine tribes. Cichorieae (21 taxa), Cardueae (11 taxa), Inuleae (six taxa), and Anthemideae (four taxa) are the leading tribes. The research included Cousinia haeckeliae, Himalaiella afghana, Pterachaenia stewartii (endemic to Afghanistan and Pakistan), and Xylanthemum macropodum (endemic to Baluchistan). Light and scanning electron microscopy were employed for the visualization of pollen photomicrographs. The data was analyzed statistically via SPSS, PAST, and Origin. Significant diagnostic qualitative and quantitative palynological traits were explored for discrimination down to the species level within the tribes. All the investigated taxa possessed radial symmetry, isopolarity, and monad form (characters for distinction at the subdivision level). The aperture types were trizonocolporate, tetrazonocolporate, and tricolporate with number position and character (NPC) formulas N3P4C5, N4P4C5, and N3P4C3. Goniotreme, peritreme, and ptychotreme types of amb were recognized. Echinate, echinate lophate, scabrate, and gemmate sculpturing were present with and without perforated surface patterns. Variations in the shapes in polar and equatorial views and lacuna shapes further assisted the separation of taxa. The observed shape classes were perprolate, prolate spheroidal, prolate, subprolate, oblate spheroidal, suboblate, and oblate. Principal component analysis, correlation, standard probability plots, and ridge line paired features plot for quantitative variables determined the positive correlation between the length and width of colpi in equatorial and polar view with polar axis and equatorial diameter and number of spines between colpi with the number of spines per pollen. The number of spines per pollen was negatively correlated with the width and length of colpi in the polar view. Multiple sample analysis of variance (ANOVA) concluded that a high statistically significant difference exists among the means of analyzed traits. The examined qualitative and quantitative palynological traits revealed noticeable variations, thus providing the source for species discrimination in Asteraceous tribes.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141157627","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Scanning electron microscopy (SEM) can reveal the ultrastructure of bacterial spores, including morphology, surface features, texture, spore damage, germination, and appendages. Understanding these features can provide a basis for adherence, how physical and environmental stressors affect spore viability, integrity, and functionality, as well as the distribution and function of surface appendages. However, the spore sample preparation method can significantly impact the SEM images' appearance, resolution, and overall quality. In this study, we compare different spore preparation methods to identify optimal approaches for preparation time, spore appearance and resolved features, including the exosporium and spore pili, for SEM imaging. We use Bacillus paranthracis as model species and evaluate the efficacy of preparation protocols using different fixation and drying methods, as well as imaging under room- and cryogenic temperatures. We compare and assess method complexity to the visibility of the spore exosporium and spore appendages across different methods. Additionally, we use Haralick texture features to quantify the differences in spore surface appearance and determine the most suitable method for preserving spore structures and surface features during SEM evaluation. The findings from this study will help establish protocols for preparing bacterial spores for SEM and facilitating accurate and reliable analysis of spores' characteristics.
{"title":"Evaluating Bacterial Spore Preparation Methods for Scanning Electron Microscopy.","authors":"Dmitry Malyshev, Cheng Choo Lee, Magnus Andersson","doi":"10.1093/mam/ozae037","DOIUrl":"10.1093/mam/ozae037","url":null,"abstract":"<p><p>Scanning electron microscopy (SEM) can reveal the ultrastructure of bacterial spores, including morphology, surface features, texture, spore damage, germination, and appendages. Understanding these features can provide a basis for adherence, how physical and environmental stressors affect spore viability, integrity, and functionality, as well as the distribution and function of surface appendages. However, the spore sample preparation method can significantly impact the SEM images' appearance, resolution, and overall quality. In this study, we compare different spore preparation methods to identify optimal approaches for preparation time, spore appearance and resolved features, including the exosporium and spore pili, for SEM imaging. We use Bacillus paranthracis as model species and evaluate the efficacy of preparation protocols using different fixation and drying methods, as well as imaging under room- and cryogenic temperatures. We compare and assess method complexity to the visibility of the spore exosporium and spore appendages across different methods. Additionally, we use Haralick texture features to quantify the differences in spore surface appearance and determine the most suitable method for preserving spore structures and surface features during SEM evaluation. The findings from this study will help establish protocols for preparing bacterial spores for SEM and facilitating accurate and reliable analysis of spores' characteristics.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140866343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mohamed A M Alsafy, Samir A A El-Gendy, Brian Atkinson, Craig J Sturrock, Basma M Kamal, Aziza Alibhai, Hanan H Abd-Elhafeez, Soha Soliman, Ahmed M Rashwan, Karam Roshdy, Catrin S Rutland
Ossa cordis, bones located within the heart trigones, are often classified as heterotopic or ectopic bones. Despite their high prevalence in cattle and some other bovids, little is known about their structure or development. Scanning electron microscopy, X-ray microtomography, gross dissections, and measurements showed the anatomical locations, prevalence, shapes, and measurements of the cardiac bones in both Egyptian Baladi cattle and Holstein-Friesians. All cattle (n = 12) had an Ossa cordis dextrum (average = 50.70 × 20.91 × 5.40 mm). Additionally, 80% Egyptian Baladi and 57% Holstein-Friesian had a smaller Ossa cordis sinistrum (average = 24.94 × 12.75 × 4.12 mm). Egyptian Baladi Ossa cordis were smaller than observed in Holstein-Friesians. Energy-dispersive X-ray analysis showed the elemental constitution (carbon, oxygen, calcium, nitrogen, phosphorus, sodium, and magnesium) of Ossa cordis and Cartilago cordis. These imaging techniques, plus four histological stains (hematoxylin and eosin, Crossman's trichrome, Alcian blue with Van Gieson, and Sirius Red) and microscopy, demonstrated osteoblasts, osteocytes, osteoclasts, astrocytes, blood vessels, bone marrow, lamellar and woven bone, cortical bone, trabeculations with pores and canaliculi, and fibrous components including collagen in the Ossa cordis dextrum and sinistrum. Hyaline cartilage and fibrocartilage (chondrocytes and cartilage matrix) were found within and surrounding the Ossa cordis. These findings were additionally compared against other cattle breeds and species.
{"title":"Novel Insights Into the Architecture of Macro and Microstructures in Cattle Ossa Cordis.","authors":"Mohamed A M Alsafy, Samir A A El-Gendy, Brian Atkinson, Craig J Sturrock, Basma M Kamal, Aziza Alibhai, Hanan H Abd-Elhafeez, Soha Soliman, Ahmed M Rashwan, Karam Roshdy, Catrin S Rutland","doi":"10.1093/mam/ozae046","DOIUrl":"10.1093/mam/ozae046","url":null,"abstract":"<p><p>Ossa cordis, bones located within the heart trigones, are often classified as heterotopic or ectopic bones. Despite their high prevalence in cattle and some other bovids, little is known about their structure or development. Scanning electron microscopy, X-ray microtomography, gross dissections, and measurements showed the anatomical locations, prevalence, shapes, and measurements of the cardiac bones in both Egyptian Baladi cattle and Holstein-Friesians. All cattle (n = 12) had an Ossa cordis dextrum (average = 50.70 × 20.91 × 5.40 mm). Additionally, 80% Egyptian Baladi and 57% Holstein-Friesian had a smaller Ossa cordis sinistrum (average = 24.94 × 12.75 × 4.12 mm). Egyptian Baladi Ossa cordis were smaller than observed in Holstein-Friesians. Energy-dispersive X-ray analysis showed the elemental constitution (carbon, oxygen, calcium, nitrogen, phosphorus, sodium, and magnesium) of Ossa cordis and Cartilago cordis. These imaging techniques, plus four histological stains (hematoxylin and eosin, Crossman's trichrome, Alcian blue with Van Gieson, and Sirius Red) and microscopy, demonstrated osteoblasts, osteocytes, osteoclasts, astrocytes, blood vessels, bone marrow, lamellar and woven bone, cortical bone, trabeculations with pores and canaliculi, and fibrous components including collagen in the Ossa cordis dextrum and sinistrum. Hyaline cartilage and fibrocartilage (chondrocytes and cartilage matrix) were found within and surrounding the Ossa cordis. These findings were additionally compared against other cattle breeds and species.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141262395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Amany Mohamed Shalaby, Shaimaa Mohamed Abdelfattah Hassan, Hanim Magdy Abdelnour, Sulaiman Mohammed Alnasser, Mohammed Alorini, Fatima A Jaber, Mohamed Ali Alabiad, Asmaa Abdullatif, Mohamed Mahmoud Abdelrahim Elshaer, Seham Ahmed Mohammed Abdel Aziz, Eman M A Abdelghany
Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease of unknown origin with limited treatment options and poor prognosis. The encouraging findings from preclinical investigations utilizing mesenchymal stem cells (MSCs) indicated that they could serve as a promising therapeutic alternative for managing chronic lung conditions, such as IPF. The objective of this study was to compare the efficiency of bone marrow-derived MSCs (BM-MSCs) versus prednisolone, the standard anti-inflammatory medication, in rats with bleomycin (BLM)-induced lung fibrosis. Four groups were created: a control group, a BLM group, a prednisolone-treated group, and a BM-MSCs-treated group. To induce lung fibrosis, 5 mg/kg of BLM was administered intratracheally. BLM significantly increased serum levels of pro-inflammatory cytokines and oxidative stress markers. The disturbed lung structure was also revealed by light and transmission electron microscopic studies. Upregulation in the immune expression of alpha-smooth muscle actin, transforming growth factor beta-1, and Bax was demonstrated. Interestingly, all findings significantly regressed on treatment with prednisolone and BM-MSCs. However, treatment with BM-MSCs showed better results than with prednisolone. In conclusion, BM-MSCs could be a promising approach for managing lung fibrosis.
{"title":"Ameliorative Potential of Bone Marrow-Derived Mesenchymal Stem Cells Versus Prednisolone in a Rat Model of Lung Fibrosis: A Histological, Immunohistochemical, and Biochemical Study.","authors":"Amany Mohamed Shalaby, Shaimaa Mohamed Abdelfattah Hassan, Hanim Magdy Abdelnour, Sulaiman Mohammed Alnasser, Mohammed Alorini, Fatima A Jaber, Mohamed Ali Alabiad, Asmaa Abdullatif, Mohamed Mahmoud Abdelrahim Elshaer, Seham Ahmed Mohammed Abdel Aziz, Eman M A Abdelghany","doi":"10.1093/mam/ozae043","DOIUrl":"10.1093/mam/ozae043","url":null,"abstract":"<p><p>Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease of unknown origin with limited treatment options and poor prognosis. The encouraging findings from preclinical investigations utilizing mesenchymal stem cells (MSCs) indicated that they could serve as a promising therapeutic alternative for managing chronic lung conditions, such as IPF. The objective of this study was to compare the efficiency of bone marrow-derived MSCs (BM-MSCs) versus prednisolone, the standard anti-inflammatory medication, in rats with bleomycin (BLM)-induced lung fibrosis. Four groups were created: a control group, a BLM group, a prednisolone-treated group, and a BM-MSCs-treated group. To induce lung fibrosis, 5 mg/kg of BLM was administered intratracheally. BLM significantly increased serum levels of pro-inflammatory cytokines and oxidative stress markers. The disturbed lung structure was also revealed by light and transmission electron microscopic studies. Upregulation in the immune expression of alpha-smooth muscle actin, transforming growth factor beta-1, and Bax was demonstrated. Interestingly, all findings significantly regressed on treatment with prednisolone and BM-MSCs. However, treatment with BM-MSCs showed better results than with prednisolone. In conclusion, BM-MSCs could be a promising approach for managing lung fibrosis.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":null,"pages":null},"PeriodicalIF":2.9,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140958447","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}