Reflection electron energy loss spectroscopy (REELS) has played a pivotal role in allowing researchers to explore the characteristics of various bulk materials. This study presents results for the low-loss region of REELS with a new cylindrical lens spectrometer integrated into a low-voltage scanning electron microscope. The operational principles and implementation of the spectrometer are explained through comparisons between electron optical simulations and experimental results. Notably, the analysis shows the ability to distinguish samples in film and bulk forms. Graphene and graphite, despite their identical elemental composition and crystalline structure, are found to have distinct energy spectra as indicated by plasmon peaks. Furthermore, the study explores the bandgap measurement of SiO2 at low-energy conditions of 2.5 keV, highlighting the proposed instrument's advantages in the measurement without the harmful effect of Cherenkov loss. Additionally, this method reaffirms the capability to measure multiple plasmon peaks from the energy spectra of bulk gold samples, thus introducing a pioneering avenue in energy spectrum measurement. Leveraging the compact size and simple experimental setup of the spectrometer for REELS, the method enables the measurement of energy spectra of both bulk- and film-formed samples under low electron energy conditions, marking a significant advancement in the field.
{"title":"A Cylindrical Lens Spectrometer with Parallel Detection for Reflection Electron Energy Loss Spectroscopy.","authors":"Junhyeok Hwang, In-Yong Park, Takashi Ogawa","doi":"10.1093/mam/ozae118","DOIUrl":"10.1093/mam/ozae118","url":null,"abstract":"<p><p>Reflection electron energy loss spectroscopy (REELS) has played a pivotal role in allowing researchers to explore the characteristics of various bulk materials. This study presents results for the low-loss region of REELS with a new cylindrical lens spectrometer integrated into a low-voltage scanning electron microscope. The operational principles and implementation of the spectrometer are explained through comparisons between electron optical simulations and experimental results. Notably, the analysis shows the ability to distinguish samples in film and bulk forms. Graphene and graphite, despite their identical elemental composition and crystalline structure, are found to have distinct energy spectra as indicated by plasmon peaks. Furthermore, the study explores the bandgap measurement of SiO2 at low-energy conditions of 2.5 keV, highlighting the proposed instrument's advantages in the measurement without the harmful effect of Cherenkov loss. Additionally, this method reaffirms the capability to measure multiple plasmon peaks from the energy spectra of bulk gold samples, thus introducing a pioneering avenue in energy spectrum measurement. Leveraging the compact size and simple experimental setup of the spectrometer for REELS, the method enables the measurement of energy spectra of both bulk- and film-formed samples under low electron energy conditions, marking a significant advancement in the field.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142730355","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}
Robert Busch, Hsu-Chih Ni, Yu-Tsun Shao, Jian-Min Zuo
We report a large-angle rocking beam electron diffraction (LARBED) technique for electron diffraction analysis. Diffraction patterns are recorded in a scanning transmission electron microscope (STEM) using a direct electron detector with large dynamical range and fast readout. We use a nanobeam for diffraction and perform the beam double rocking by synchronizing the detector with the STEM scan coils for the recording. Using this approach, large-angle convergent beam electron diffraction (LACBED) patterns of different reflections are obtained simultaneously. By using a nanobeam, instead of a focused beam, the LARBED technique can be applied to beam-sensitive crystals as well as crystals with large unit cells. This paper describes the implementation of LARBED and evaluates the performance using silicon and gadolinium gallium garnet crystals as test samples. We demonstrate that our method provides an effective and robust way for recording LARBED patterns and paves the way for quantitative electron diffraction of large unit cell and beam-sensitive crystals.
{"title":"Large-Angle Rocking Beam Electron Diffraction of Large Unit Cell Crystals Using Direct Electron Detector.","authors":"Robert Busch, Hsu-Chih Ni, Yu-Tsun Shao, Jian-Min Zuo","doi":"10.1093/mam/ozae088","DOIUrl":"10.1093/mam/ozae088","url":null,"abstract":"<p><p>We report a large-angle rocking beam electron diffraction (LARBED) technique for electron diffraction analysis. Diffraction patterns are recorded in a scanning transmission electron microscope (STEM) using a direct electron detector with large dynamical range and fast readout. We use a nanobeam for diffraction and perform the beam double rocking by synchronizing the detector with the STEM scan coils for the recording. Using this approach, large-angle convergent beam electron diffraction (LACBED) patterns of different reflections are obtained simultaneously. By using a nanobeam, instead of a focused beam, the LARBED technique can be applied to beam-sensitive crystals as well as crystals with large unit cells. This paper describes the implementation of LARBED and evaluates the performance using silicon and gadolinium gallium garnet crystals as test samples. We demonstrate that our method provides an effective and robust way for recording LARBED patterns and paves the way for quantitative electron diffraction of large unit cell and beam-sensitive crystals.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142365821","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}
Paclitaxel, a chemotherapeutic drug, induces sensorimotor peripheral neuropathy. Carvedilol, a nonselective β-adrenoreceptor blocker, has been shown to exert antioxidant activity. Platelet-rich plasma (PRP) has supra-physiological levels of growth factors (GFs), enhances biosynthesis of antioxidant enzymes, and suppresses oxidative stress. This study compared the ameliorative effects of carvedilol and PRP on paclitaxel-induced femoral neuropathy. Eighty-eight adult male albino rats were equally randomized into four groups: group I served as the control; group II received paclitaxel (16 mg/kg intraperitoneally, weekly); group III received carvedilol (10 mg/kg daily, orally) concomitant with paclitaxel; and group IV received PRP (0.5 mL/kg subcutaneously, twice weekly) concomitant with paclitaxel. After 5 weeks, femoral nerve conduction velocity was measured, and blood samples were collected to assess catalase and superoxide dismutase levels. All animals were sacrificed, and gene expression of miR-21 was quantified. Tissue sections were stained with hematoxylin and eosin and toluidine blue. Then, the ultrathin sections were examined by transmission electron microscopy. Both carvedilol and PRP reversed paclitaxel-induced changes in the peripheral nerve, but PRP demonstrated a stronger antioxidant effect and a more pronounced presence of GFs, as evidenced by electron microscopy. PRP may represent a promising therapeutic approach for paclitaxel-induced neuropathy.
{"title":"Ameliorative Potential of Carvedilol Versus Platelet-Rich Plasma Against Paclitaxel-Induced Femoral Neuropathy in Wistar Rats: A Light and Electron Microscopic Study.","authors":"Ereny Fekry, George Nagi Refaat, Sara Adel Hosny","doi":"10.1093/mam/ozaf002","DOIUrl":"10.1093/mam/ozaf002","url":null,"abstract":"<p><p>Paclitaxel, a chemotherapeutic drug, induces sensorimotor peripheral neuropathy. Carvedilol, a nonselective β-adrenoreceptor blocker, has been shown to exert antioxidant activity. Platelet-rich plasma (PRP) has supra-physiological levels of growth factors (GFs), enhances biosynthesis of antioxidant enzymes, and suppresses oxidative stress. This study compared the ameliorative effects of carvedilol and PRP on paclitaxel-induced femoral neuropathy. Eighty-eight adult male albino rats were equally randomized into four groups: group I served as the control; group II received paclitaxel (16 mg/kg intraperitoneally, weekly); group III received carvedilol (10 mg/kg daily, orally) concomitant with paclitaxel; and group IV received PRP (0.5 mL/kg subcutaneously, twice weekly) concomitant with paclitaxel. After 5 weeks, femoral nerve conduction velocity was measured, and blood samples were collected to assess catalase and superoxide dismutase levels. All animals were sacrificed, and gene expression of miR-21 was quantified. Tissue sections were stained with hematoxylin and eosin and toluidine blue. Then, the ultrathin sections were examined by transmission electron microscopy. Both carvedilol and PRP reversed paclitaxel-induced changes in the peripheral nerve, but PRP demonstrated a stronger antioxidant effect and a more pronounced presence of GFs, as evidenced by electron microscopy. PRP may represent a promising therapeutic approach for paclitaxel-induced neuropathy.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143189909","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}
Ellis Kennedy, Emily Hollingworth, Alejandro Ceballos, Daisy O'Mahoney, Colin Ophus, Frances Hellman, Mary Scott
Amorphous thin films grown by magnetron co-sputtering exhibit changes in atomic structure with varying growth and annealing temperatures. Structural variations influence the bulk properties of the films. Scanning nanodiffraction performed in a transmission electron microscope (TEM) is applied to amorphous Tb17Co83 (a-Tb-Co) films deposited over a range of temperatures to measure relative changes in medium-range ordering (MRO). These measurements reveal an increase in MRO with higher growth temperatures and a decrease in MRO with higher annealing temperatures. The trend in MRO indicates a relationship between the growth conditions and local atomic ordering. By tilting select films, the TEM measures variations in the local atomic structure as a function of orientation within the films. The findings support claims that preferential ordering along the growth direction results from temperature-mediated adatom configurations during deposition, and that oriented MRO correlates with increased structural anisotropy, explaining the strong growth-induced perpendicular magnetic anisotropy found in rare earth-transition metal films. Beyond magnetic films, we propose the tilted FEM workflow as a method of extracting anisotropic structural information in a variety of amorphous materials with directionally dependent bulk properties, such as films with inherent bonding asymmetry grown by physical vapor deposition.
{"title":"Exploring Structural Anisotropy in Amorphous Tb-Co via Changes in Medium-Range Ordering.","authors":"Ellis Kennedy, Emily Hollingworth, Alejandro Ceballos, Daisy O'Mahoney, Colin Ophus, Frances Hellman, Mary Scott","doi":"10.1093/mam/ozae113","DOIUrl":"10.1093/mam/ozae113","url":null,"abstract":"<p><p>Amorphous thin films grown by magnetron co-sputtering exhibit changes in atomic structure with varying growth and annealing temperatures. Structural variations influence the bulk properties of the films. Scanning nanodiffraction performed in a transmission electron microscope (TEM) is applied to amorphous Tb17Co83 (a-Tb-Co) films deposited over a range of temperatures to measure relative changes in medium-range ordering (MRO). These measurements reveal an increase in MRO with higher growth temperatures and a decrease in MRO with higher annealing temperatures. The trend in MRO indicates a relationship between the growth conditions and local atomic ordering. By tilting select films, the TEM measures variations in the local atomic structure as a function of orientation within the films. The findings support claims that preferential ordering along the growth direction results from temperature-mediated adatom configurations during deposition, and that oriented MRO correlates with increased structural anisotropy, explaining the strong growth-induced perpendicular magnetic anisotropy found in rare earth-transition metal films. Beyond magnetic films, we propose the tilted FEM workflow as a method of extracting anisotropic structural information in a variety of amorphous materials with directionally dependent bulk properties, such as films with inherent bonding asymmetry grown by physical vapor deposition.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142623881","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}
Connor J Parker, Kathryn M Peruski, Samantha K Cary
The production of plutonium-238 through irradiation of neptunium-237 (237Np) target materials for the use in radioisotope thermoelectric generators is paramount for continued deep space exploration. This work employs scanning electron microscopy to analyze 237Np materials coupled with a well-developed image analysis framework (Morphological Analysis for Material Attribution, or MAMA) to determine the degree of micron-scale homogeneity in the materials. This work demonstrated how the quantification of particle characteristics can validate production materials and affirm the qualitative similarities observed in micrographs. The 237Np oxide particle analysis determined that the materials from five production runs were quantitatively homogenous (significant at α = 0.05) in particle area, circularity, equivalent circular diameter, and ellipse aspect ratio, with two of the sampling dates having statistically significant different means for one of the four characteristics. These metrics not only confirm general homogeneity of the material but also expand the application of MAMA workflows to 237Np materials, demonstrating the utility of MAMA analysis for a wider breadth of nuclear materials than previously reported. In the open literature, this study is the first time that these microanalytical techniques were applied to 237Np materials to this degree.
通过辐照镎237(237Np)目标材料来生产钚238,并将其用于放射性同位素热电发电机,这对于继续进行深空探测至关重要。这项工作利用扫描电子显微镜分析 237Np 材料,并结合完善的图像分析框架(材料归属形态分析,或 MAMA)来确定材料的微米级均匀程度。这项工作展示了颗粒特征的量化如何验证生产材料并确认显微照片中观察到的定性相似性。237Np 氧化物颗粒分析表明,来自五次生产运行的材料在颗粒面积、圆度、等效圆直径和椭圆长宽比方面具有定量同质性(α = 0.05 时显著),其中两个取样日期的四个特征之一的平均值差异具有统计学意义。这些指标不仅证实了材料的总体均匀性,还将 MAMA 工作流程的应用扩展到了 237Np 材料,证明了 MAMA 分析在比以往报告更广泛的核材料中的实用性。在公开文献中,这项研究是首次将这些微分析技术应用于 237Np 材料。
{"title":"Quantitative Particle Analysis of Neptunium-237 Oxides: Optimization of MAMA Analysis for Modified Direct Denitration Products.","authors":"Connor J Parker, Kathryn M Peruski, Samantha K Cary","doi":"10.1093/mam/ozae112","DOIUrl":"10.1093/mam/ozae112","url":null,"abstract":"<p><p>The production of plutonium-238 through irradiation of neptunium-237 (237Np) target materials for the use in radioisotope thermoelectric generators is paramount for continued deep space exploration. This work employs scanning electron microscopy to analyze 237Np materials coupled with a well-developed image analysis framework (Morphological Analysis for Material Attribution, or MAMA) to determine the degree of micron-scale homogeneity in the materials. This work demonstrated how the quantification of particle characteristics can validate production materials and affirm the qualitative similarities observed in micrographs. The 237Np oxide particle analysis determined that the materials from five production runs were quantitatively homogenous (significant at α = 0.05) in particle area, circularity, equivalent circular diameter, and ellipse aspect ratio, with two of the sampling dates having statistically significant different means for one of the four characteristics. These metrics not only confirm general homogeneity of the material but also expand the application of MAMA workflows to 237Np materials, demonstrating the utility of MAMA analysis for a wider breadth of nuclear materials than previously reported. In the open literature, this study is the first time that these microanalytical techniques were applied to 237Np materials to this degree.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142623900","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}
Barleria is a palaeotropical genus of herbs, shrubs, and rarely climbers or trees. We investigated the karyotypes and male meiosis of 12 and 13 species, respectively, for the first time. Mitotic metaphases revealed two chromosome counts, 2n = 40 and 2n = 44. Chromosomes had median (m), submedian (sm), and subterminal (st) region centromeres. The total haploid chromosome length (TCL) ranged from 78.95 µm (Barleria sahyadrica) to 37.80 µm (B. nitida). Dispersion index differentiated the species into two groups, one with lower (3.40-4.79) and the other with higher (6.63-12.87) values. Principal component analysis based on six karyological parameters, namely base number (x), 2n, TCL, coefficient of variation of chromosome length, coefficient of variation of centromeric index, and mean centromeric asymmetry, exhibited three clusters. Cluster I included species of the subgenus Barleria. Cluster III had species of the subgenus Prionitis section Somalia. Cluster II comprised species of the subgenus Barleria and the subgenus Prionitis section Prionitis (B. sahyadrica). Pollen grains were oblate spheroidal or distinctly three-lobed, tri-brevicolporate with honey-combed tectum. Our analyses revealed karyological relationships among the investigated species and also provide raw data to breeders interested in horticultural applications of Barleria for accomplishing interspecific hybridization.
{"title":"Karyotype, Male Meiosis, and Pollen Features of Barleria (Acanthaceae): A Wild Ornamental From India.","authors":"Suraj S Patil, Manoj M Lekhak","doi":"10.1093/mam/ozae117","DOIUrl":"10.1093/mam/ozae117","url":null,"abstract":"<p><p>Barleria is a palaeotropical genus of herbs, shrubs, and rarely climbers or trees. We investigated the karyotypes and male meiosis of 12 and 13 species, respectively, for the first time. Mitotic metaphases revealed two chromosome counts, 2n = 40 and 2n = 44. Chromosomes had median (m), submedian (sm), and subterminal (st) region centromeres. The total haploid chromosome length (TCL) ranged from 78.95 µm (Barleria sahyadrica) to 37.80 µm (B. nitida). Dispersion index differentiated the species into two groups, one with lower (3.40-4.79) and the other with higher (6.63-12.87) values. Principal component analysis based on six karyological parameters, namely base number (x), 2n, TCL, coefficient of variation of chromosome length, coefficient of variation of centromeric index, and mean centromeric asymmetry, exhibited three clusters. Cluster I included species of the subgenus Barleria. Cluster III had species of the subgenus Prionitis section Somalia. Cluster II comprised species of the subgenus Barleria and the subgenus Prionitis section Prionitis (B. sahyadrica). Pollen grains were oblate spheroidal or distinctly three-lobed, tri-brevicolporate with honey-combed tectum. Our analyses revealed karyological relationships among the investigated species and also provide raw data to breeders interested in horticultural applications of Barleria for accomplishing interspecific hybridization.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142739540","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}
Anna Scheid, Isha, Arvind Kumar Yogi, Masahiko Isobe, Birgit Bußmann, Tobias Heil, Peter A van Aken
In the field of quantum materials, understanding anomalous behavior under charge degrees of freedom through bond formation is of fundamental importance, with two key concepts: Dimerization and charge order at different cation sites. The coexistence of both dimerization and charge ordering is unusually found in NaRu2O4, even in its metallic state at room temperature. Our work unveils the origin of the interplay of these effects within metallic single-crystalline NaRu2O4. Employing advanced transmission electron microscopy techniques, we probe the lattice order of NaRu2O4 as a function of temperature and provide direct microscopic evidence of a Peierls-type transition. This transition is accompanied by a pronounced dimerization of the ruthenium chains, resulting in a distinctive twofold superstructure along the b axis below the critical transition temperature of ∼535 K, coinciding with a charge order. In situ heating experiments confirm the reversibility of this first-order phase transition, and periodic lattice displacement maps depict atomic-scale displacements linked to dimerization.
{"title":"Direct Evidence of Anomalous Peierls Transition-Induced Charge Density Wave Order at Room Temperature in Metallic NaRu2O4.","authors":"Anna Scheid, Isha, Arvind Kumar Yogi, Masahiko Isobe, Birgit Bußmann, Tobias Heil, Peter A van Aken","doi":"10.1093/mam/ozae129","DOIUrl":"10.1093/mam/ozae129","url":null,"abstract":"<p><p>In the field of quantum materials, understanding anomalous behavior under charge degrees of freedom through bond formation is of fundamental importance, with two key concepts: Dimerization and charge order at different cation sites. The coexistence of both dimerization and charge ordering is unusually found in NaRu2O4, even in its metallic state at room temperature. Our work unveils the origin of the interplay of these effects within metallic single-crystalline NaRu2O4. Employing advanced transmission electron microscopy techniques, we probe the lattice order of NaRu2O4 as a function of temperature and provide direct microscopic evidence of a Peierls-type transition. This transition is accompanied by a pronounced dimerization of the ruthenium chains, resulting in a distinctive twofold superstructure along the b axis below the critical transition temperature of ∼535 K, coinciding with a charge order. In situ heating experiments confirm the reversibility of this first-order phase transition, and periodic lattice displacement maps depict atomic-scale displacements linked to dimerization.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142979159","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}
There is still room for improvement in the isolation and purification techniques for extracellular vesicles (EVs), particularly in the separation of exosomes (small EVs) from other membrane vesicles such as microvesicles and apoptotic bodies. Furthermore, it is crucial to establish preparation methods that preserve the intrinsic properties of EVs in this context. In this study, we focus on the isolation and preparation of small EVs, exosomes, from the culture supernatant of a human cell line. We discuss the sequential use of regenerated cellulose membranes with different molecular weight cutoffs, based on direct evaluation by transmission electron microscopy, and examine the challenges of characterizing biological membrane vesicles, small EVs, identified during this process.
{"title":"Preparation of Small Extracellular Vesicles Using Sequential Ultrafiltration with Regenerated Cellulose Membranes of Different Molecular Weight Cutoffs: A Study of Morphology and Size by Electron Microscopy.","authors":"Noriyuki Ishii, Hiroaki Tateno","doi":"10.1093/mam/ozae133","DOIUrl":"10.1093/mam/ozae133","url":null,"abstract":"<p><p>There is still room for improvement in the isolation and purification techniques for extracellular vesicles (EVs), particularly in the separation of exosomes (small EVs) from other membrane vesicles such as microvesicles and apoptotic bodies. Furthermore, it is crucial to establish preparation methods that preserve the intrinsic properties of EVs in this context. In this study, we focus on the isolation and preparation of small EVs, exosomes, from the culture supernatant of a human cell line. We discuss the sequential use of regenerated cellulose membranes with different molecular weight cutoffs, based on direct evaluation by transmission electron microscopy, and examine the challenges of characterizing biological membrane vesicles, small EVs, identified during this process.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142979178","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}
Ian MacLaren, Andrew T Fraser, Matthew R Lipsett, Colin Ophus
A new method for dark field imaging is introduced, which uses scanned electron diffraction (or 4DSTEM-4-dimensional scanning transmission electron microscopy) datasets as its input. Instead of working on simple summation of intensity, it works on a sparse representation of the diffraction patterns in terms of a list of their diffraction peaks. This is tested on a thin perovskite film containing structural ordering resulting in additional superlattice spots that reveal details of domain structures, and is shown to give much better selectivity and contrast than conventional virtual dark field imaging. It is also shown to work well in polycrystalline aggregates of CuO nanoparticles. In view of the higher contrast and selectivity, and the complete exclusion of diffuse scattering from the image formation, it is expected to be of significant benefit for characterization of a wide variety of crystalline materials.
{"title":"Digital Dark Field-Higher Contrast and Greater Specificity Dark Field Imaging Using a 4DSTEM Approach.","authors":"Ian MacLaren, Andrew T Fraser, Matthew R Lipsett, Colin Ophus","doi":"10.1093/mam/ozae104","DOIUrl":"10.1093/mam/ozae104","url":null,"abstract":"<p><p>A new method for dark field imaging is introduced, which uses scanned electron diffraction (or 4DSTEM-4-dimensional scanning transmission electron microscopy) datasets as its input. Instead of working on simple summation of intensity, it works on a sparse representation of the diffraction patterns in terms of a list of their diffraction peaks. This is tested on a thin perovskite film containing structural ordering resulting in additional superlattice spots that reveal details of domain structures, and is shown to give much better selectivity and contrast than conventional virtual dark field imaging. It is also shown to work well in polycrystalline aggregates of CuO nanoparticles. In view of the higher contrast and selectivity, and the complete exclusion of diffuse scattering from the image formation, it is expected to be of significant benefit for characterization of a wide variety of crystalline materials.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142623871","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}
Marcel Schloz, Thomas C Pekin, Hamish G Brown, Dana O Byrne, Bryan D Esser, Emmanuel Terzoudis-Lumsden, Takashi Taniguchi, Kenji Watanabe, Scott D Findlay, Benedikt Haas, Jim Ciston, Christoph T Koch
A detailed analysis of ptychography for three-dimensional (3D) phase reconstructions of thick specimens is performed. We introduce multi-focus ptychography, which incorporates a 4D-STEM defocus series to enhance the quality of 3D reconstructions along the beam direction through a higher overdetermination ratio. This method is compared with established multi-slice ptychography techniques, such as conventional ptychography, regularized ptychography, and multi-mode ptychography. Additionally, we contrast multi-focus ptychography with an alternative method that uses virtual optical sectioning through a reconstructed scattering matrix (S-matrix), which offers more precise 3D structure information compared to conventional ptychography. Our findings from multiple 3D reconstructions based on simulated and experimental data demonstrate that multi-focus ptychography surpasses other techniques, particularly in accurately reconstructing the surfaces and interface regions of thick specimens.
{"title":"Improved Three-Dimensional Reconstructions in Electron Ptychography through Defocus Series Measurements.","authors":"Marcel Schloz, Thomas C Pekin, Hamish G Brown, Dana O Byrne, Bryan D Esser, Emmanuel Terzoudis-Lumsden, Takashi Taniguchi, Kenji Watanabe, Scott D Findlay, Benedikt Haas, Jim Ciston, Christoph T Koch","doi":"10.1093/mam/ozae110","DOIUrl":"10.1093/mam/ozae110","url":null,"abstract":"<p><p>A detailed analysis of ptychography for three-dimensional (3D) phase reconstructions of thick specimens is performed. We introduce multi-focus ptychography, which incorporates a 4D-STEM defocus series to enhance the quality of 3D reconstructions along the beam direction through a higher overdetermination ratio. This method is compared with established multi-slice ptychography techniques, such as conventional ptychography, regularized ptychography, and multi-mode ptychography. Additionally, we contrast multi-focus ptychography with an alternative method that uses virtual optical sectioning through a reconstructed scattering matrix (S-matrix), which offers more precise 3D structure information compared to conventional ptychography. Our findings from multiple 3D reconstructions based on simulated and experimental data demonstrate that multi-focus ptychography surpasses other techniques, particularly in accurately reconstructing the surfaces and interface regions of thick specimens.</p>","PeriodicalId":18625,"journal":{"name":"Microscopy and Microanalysis","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142668369","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}