Ju Ran Byeon, Eun Mi Song, Yang Hee Joo, A Reum Choe, Yehyun Park, Chung Hyun Tae, Chang Mo Moon, Seong-Eun Kim, Hye-Kyung Jung, Ki-Nam Shim, Sung-Ae Jung
{"title":"Exploring the Morphologic Characteristics of Intraperitoneally Injected Three-Dimensional-Cultured Tonsil-Derived Mesenchymal Stem Cells in a Murine Colitis Model Using Electron Microscopy.","authors":"Ju Ran Byeon, Eun Mi Song, Yang Hee Joo, A Reum Choe, Yehyun Park, Chung Hyun Tae, Chang Mo Moon, Seong-Eun Kim, Hye-Kyung Jung, Ki-Nam Shim, Sung-Ae Jung","doi":"10.1093/mam/ozae135","DOIUrl":"https://doi.org/10.1093/mam/ozae135","url":null,"abstract":"<p><p>Three-dimensional (3D)-cultured tonsil-derived mesenchymal stem cells (TMSCs) show high therapeutic efficacy in murine colitis. We evaluated the in vivo localization and formation of 3D-TMSCs using electron microscopy. Mice with dextran sulfate sodium-induced chronic colitis received intraperitoneal injections of 3D-TMSCs. The formation and localization of TMSC aggregates in the peritoneal cavity were assessed by immunofluorescence and electron microscopy. In mice treated with 3D-TMSCs, white spherical aggregates attached to the peritoneal cavity were observed. Immunofluorescence staining revealed the co-localization of human-origin TMSCs and mouse immune cells within these aggregates. The presence of human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) confirmed the presence of human-origin TMSCs in the aggregates. Electron microscopy revealed that 3D-TMSCs were covered with mouse immune cells. The presence of pseudopodia and microvilli facilitated cell-to-cell connections, indicating a complex integrated structure rather than a mere collection of TMSCs. After 15 days, rough endoplasmic reticulum and mitochondria were identified within the cytoplasm of the TMSCs. Prominent autophagosomes and extracellular vesicles were observed within the intraperitoneal 3D-TMSC aggregates. These findings underscore the viability and paracrine effects of 3D-TMSCs and support their potential as an advanced therapeutic option for treating inflammatory bowel disease.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145131314","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}
Rebecca C Leghziel, Lia Addadi, Assaf Gal, Lothar Houben
{"title":"Solving the Crystal Architecture of Coccoliths Using 4D-STEM.","authors":"Rebecca C Leghziel, Lia Addadi, Assaf Gal, Lothar Houben","doi":"10.1093/mam/ozaf087","DOIUrl":"https://doi.org/10.1093/mam/ozaf087","url":null,"abstract":"<p><p>Marine phytoplankton form functional biominerals with intricate morphologies and architectures. Coccolithophores occupy a special position among these organisms because of their production of intricate calcite scales, called coccoliths. Although coccolith morphologies differ across different species, crystals are organized around an organic matrix systematically to form an arrangement of astounding symmetry. We demonstrate the opportunities emerging from four-dimensional scanning transmission electron microscopy (4D-STEM), to spatially solve the crystallography of such biominerals. Through the development of a computational pipeline, which automatically solves the orientation at image pixels corresponding to crystals, we can map the orientation of the entangled and overlapping crystalline building blocks composing the coccolith. The present work exemplifies how parallel real space and diffraction space recordings can facilitate and improve the throughput of deciphering the complex network of biomineral superstructures.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145244513","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: Comprehensive Comparison Between STEM-HAADF and TEM Bright-field Mode for Imaging Resin Embedded Biological Samples.","authors":"","doi":"10.1093/mam/ozaf088","DOIUrl":"https://doi.org/10.1093/mam/ozaf088","url":null,"abstract":"","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145301568","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}
Christoph Flathmann, Ulrich Ross, Jürgen Belz, Andreas Beyer, Kerstin Volz, Michael Seibt, Tobias Meyer
{"title":"Sequential Tilting 4D-STEM for Improved Momentum-Resolved STEM Field Mapping.","authors":"Christoph Flathmann, Ulrich Ross, Jürgen Belz, Andreas Beyer, Kerstin Volz, Michael Seibt, Tobias Meyer","doi":"10.1093/mam/ozaf086","DOIUrl":"https://doi.org/10.1093/mam/ozaf086","url":null,"abstract":"<p><p>Momentum-resolved scanning transmission electron microscopy (MRSTEM) is a powerful phase-contrast technique that can map lateral magnetic and electric fields ranging from the micrometer to the subatomic scale. Resolving fields ranging from a few nanometers to a few hundred nanometers, as well as across material interfaces, is particularly important since these fields often determine the functional properties of devices. However, it is also challenging since they are orders of magnitude smaller than atomic electric fields. Thus, subtle changes in diffraction conditions lead to significant changes in the measured MRSTEM signal. One established approach to partially overcome this problem is precession electron diffraction, in which the incident electron beam is continuously precessed while precession-averaged diffraction patterns are acquired. Here, we present an alternative approach in which we sequentially tilt the incident electron beam and record a full diffraction pattern for each tilt and spatial position. This approach requires no hardware modification of the instrument and enables the use of arbitrary beam tilt patterns that can be optimized for specific applications. Furthermore, recording diffraction patterns for every beam tilt allows access to additional information. In this work, we use this information to create virtual large-angle convergent beam electron diffraction patterns to assess MRSTEM data quality and improve field measurements by applying different data analysis methods beyond simple averaging. The presented data acquisition concept can readily be applied to other 4D-STEM applications.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145308632","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":"Efferocytosis-Driven M2 Macrophage Impairs Fibrotic Encapsulation and Promotes Echinococcus multilocularis Growth in Cotton Rats (Sigmodon hispidus).","authors":"Maru Manabe, Teppei Nakamura, Keisuke Sato, Naoki Hayashi, Hirokazu Kouguchi, Ryo Nakao, Masahito Hidaka, Hiroyuki Matsuyama, Nariaki Nonaka, Masami Morimatsu","doi":"10.1093/mam/ozaf082","DOIUrl":"10.1093/mam/ozaf082","url":null,"abstract":"<p><p>Alveolar echinococcosis, caused by Echinococcus multilocularis, exhibits significant species-dependent susceptibility. This study compared the early hepatic tissue responses to E. multilocularis in highly susceptible cotton rats (Sigmodon hispidus) and laboratory mice (DBA/2 and AKR/N). Following oral administration of E. multilocularis eggs, cotton rats developed a greater number of hepatic lesions within 2 weeks, whereas mice required 4 weeks to develop smaller lesions. Histopathology revealed accelerated multilocular cyst formation in cotton rats. Unlike mice, which formed dense collagenous layers isolating cysts, cotton rats lacked adventitial layers despite similar fibrotic thickness. Immunohistochemistry revealed abundant CD206+ macrophages at cyst peripheries in cotton rats, engaging in efferocytosis of apoptotic neutrophils with expression of TGF-β, galectin-3, and VEGF. Efferocytic macrophages expressed collagen-degrading enzymes (cathepsin K and MMP9) and the growth factor FGF2. These findings suggest that efferocytosis by neutrophils drives macrophages toward an anti-inflammatory M2 phenotype, leading to immune evasion, ineffective fibrotic encapsulation, and parasitic growth. Given the wide distribution of cotton rats in the Americas and the expanding range of E. multilocularis, their hypersusceptibility raises significant public health concerns as rodents could serve as an intermediate host. These insights may inform new strategies for host-parasite interactions and the control of alveolar echinococcosis.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144961091","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}
Fernando Camino, Byeongjun Gil, Armando Rúa, Meng Li, Dmitri N Zakharov, Kim Kisslinger, Myung-Geun Han, Daniel C Hayes, Juan Alban, Rakesh Agrawal, Miyoung Kim, Yimei Zhu, Judith C Yang
{"title":"Enabling the Acquisition of Electron Beam-Induced Current (EBIC) Images in Conventional SEM and STEM Instruments.","authors":"Fernando Camino, Byeongjun Gil, Armando Rúa, Meng Li, Dmitri N Zakharov, Kim Kisslinger, Myung-Geun Han, Daniel C Hayes, Juan Alban, Rakesh Agrawal, Miyoung Kim, Yimei Zhu, Judith C Yang","doi":"10.1093/mam/ozaf080","DOIUrl":"https://doi.org/10.1093/mam/ozaf080","url":null,"abstract":"<p><p>Electron beam-induced current (EBIC) imaging is a well-established scanning electron microscope (SEM) technique used to analyze the behavior of microelectronic devices including solar cells. Recently, the application of EBIC imaging in an aberration-corrected scanning transmission electron microscope (STEM) has been demonstrated and offers great potential for the in situ study of electronic materials, correlating charge transport properties to atomic structural and elemental information. This work presents two ways to implement EBIC imaging in conventional SEM and STEM systems: one relying on the instrument's inherent scanning and imaging electronics and the other involving third-party systems usually available in electron microscopes. The implementation of lock-in EBIC in systems equipped with a fast beam blanker is also described. In addition, this work shows and discusses the different mechanisms at play in EBIC imaging and their dependence on beam energy, sample impedance, and electrical measurement configuration, providing researchers with the basic information needed to apply the technique to their research.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 5","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145131273","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}
Abigail Carbone, Robert Sinclair, Reinhold H Dauskardt
{"title":"Advancing Cross-Sectional Scanning Electron Microscopy of Perovskite Solar Cells.","authors":"Abigail Carbone, Robert Sinclair, Reinhold H Dauskardt","doi":"10.1093/mam/ozaf067","DOIUrl":"https://doi.org/10.1093/mam/ozaf067","url":null,"abstract":"<p><p>Organic-inorganic perovskites are an emerging class of photovoltaic materials. Despite achieving power conversion efficiencies surpassing 26%, the challenge of perovskite stability including degradation during exposure to operational conditions such as light, heat, humidity, water, oxygen, and electric fields is well known. Related, perovskite instability has limited high-resolution electron imaging and characterization techniques that can be used for understanding degradation mechanisms. Here, we demonstrate perovskite device cross-section preparation using mechanical polishing in a water-free environment with cryogenic Ar ion milling. Scanning electron microscopy was then used in both backscattered electron and secondary electron imaging modes to obtain information about layer structure, grain aggregate structure, and compositional heterogeneity. Monte Carlo CASINO simulations inform optimum beam conditions and image acquisition parameters and the effects of accelerating voltage, dwell times, and frame averaging for practical image acquisition are reported.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 4","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144690926","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":"Multi-laboratory Validation of Join of the Five Dyes Revealing Collagenous Tissue (JFRL) Staining for Bone Histology Across Animal Species and Bone Implant Materials.","authors":"Erika Tsuji, Kanako Sumi, Natsuko Tsuzuki, Daisuke Kondoh, Masashi Tsujio, Marina Hosotani, Takashi Namba, Shoichi Wakitani, Osamu Ichii, Ko Nakanishi, Teppei Nakamura","doi":"10.1093/mam/ozaf058","DOIUrl":"https://doi.org/10.1093/mam/ozaf058","url":null,"abstract":"<p><p>Histological staining is essential for understanding bone structure and pathology; however, variations in decalcification agents can compromise reproducibility. We have previously developed a novel osteochondral staining method, Join of the Five dyes Revealing coLlagenous tissue (JFRL) staining, that is independent of the decalcification method. To promote its widespread adoption, this study confirms the robustness of JFRL staining through intra- and inter-laboratory validation. JFRL staining demonstrated consistent patterns across different manufacturers and facilities, with proper dehydration steps being crucial for optimal results. We applied JFRL staining to diverse vertebrate species prepared under various fixation and decalcification conditions to effectively visualize species-specific bone structures, including distinct osteoid and mineralized bone features from fish to large mammals. Furthermore, JFRL staining proved useful in evaluating bone biomaterials within defect models and clearly depicts the complex architecture of bone-healing processes and material integration. The staining qualitatively distinguished osteoid, mineralized bone, hyaline cartilage, and bone cells of different colors across all applications. These findings establish JFRL staining as a robust and versatile method for bone histology. Future studies focusing on quantitative assessment and pathological applications will prove that JFRL staining presents a reliable tool for both basic research and clinical diagnostics of bone disorders.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 4","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144637613","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}
Kyvia L C Costa, Fernanda C R Dias, Fabiana C S A Melo, Marcos L M Gomes, Sérgio L P Matta
{"title":"Deleterious Effects of Lead Acetate Intake on the Spermatogenic Process in Adult Wistar Rats.","authors":"Kyvia L C Costa, Fernanda C R Dias, Fabiana C S A Melo, Marcos L M Gomes, Sérgio L P Matta","doi":"10.1093/mam/ozaf074","DOIUrl":"10.1093/mam/ozaf074","url":null,"abstract":"<p><p>Lead is a widespread environmental pollutant that affects biological systems, particularly the male reproductive system. This study evaluated the effects of subchronic lead exposure on the testicular parenchyma of adult Wistar rats. Animals were divided into five groups: one control and four treated with increasing doses of lead acetate (16, 32, 64, and 128 mg/kg/day). Testicular tissues were analyzed using light and transmission electron microscopy, along with measurements of testicular lead concentration and antioxidant enzyme activity (SOD and CAT). Histopathological alterations included vacuolization, germ cell desquamation, apoptotic bodies, lipid droplets, and blood-testis barrier rupture. A dose-dependent reduction in seminiferous epithelium height and germ cell population was observed, along with an enlarged tubular lumen. These structural changes resulted in decreased sperm production and sperm reserves, particularly at higher lead doses. Additionally, lead exposure significantly reduced the activity of SOD and CAT enzymes, indicating oxidative stress. In conclusion, subchronic lead exposure disrupts testicular structure and function by inducing oxidative damage, leading to impaired spermatogenesis and fertility.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144883188","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}
Aurélie Dehlinger, Valentina Alberini, Vladimir Usatikov, Céline Dyhring, Sarah Jung, Jonas Grage, Daniel Grötzsch, Johannes Tümmler, Stefan Rehbein, Holger Stiel, Birgit Kanngießer, Christian Seim
{"title":"Laboratory-Based Soft X-Ray Tomography: A Novel Laser Design, Source Monitoring, and Data-Processing Workflow.","authors":"Aurélie Dehlinger, Valentina Alberini, Vladimir Usatikov, Céline Dyhring, Sarah Jung, Jonas Grage, Daniel Grötzsch, Johannes Tümmler, Stefan Rehbein, Holger Stiel, Birgit Kanngießer, Christian Seim","doi":"10.1093/mam/ozaf073","DOIUrl":"10.1093/mam/ozaf073","url":null,"abstract":"<p><p>Method development for laboratory-based X-ray microscopes operating in the water-window range invariably involves the development of the X-ray source as well. This paper presents major upgrades to the laboratory soft X-ray microscope (L-TXM) plasma chamber and data analysis protocol. Characterization of the laser-plasma source demonstrates improved performance, while a proof-of-principle tomogram of a diatom showcases a robust data treatment protocol and the system's capabilities for three-dimensional imaging and segmentation. These developments mark significant progress toward making L-TXM a more robust and user-friendly tool for soft X-ray microscopy applications.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":"31 4","pages":""},"PeriodicalIF":3.0,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144883192","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}