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TEM and STEM-EDS evaluation of metal nanoparticle encapsulation in GroEL/GroES complexes according to the reaction mechanism of chaperonin 根据伴侣蛋白的反应机理对GroEL/GroES复合物中金属纳米粒子包封的TEM和STEM-EDS评估
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa064
Hiromi Yoda;Ayumi Koike-Takeshita
{"title":"TEM and STEM-EDS evaluation of metal nanoparticle encapsulation in GroEL/GroES complexes according to the reaction mechanism of chaperonin","authors":"Hiromi Yoda;Ayumi Koike-Takeshita","doi":"10.1093/jmicro/dfaa064","DOIUrl":"10.1093/jmicro/dfaa064","url":null,"abstract":"Escherichia coli chaperonin GroEL, which is a large cylindrical protein complex comprising two heptameric rings with cavities of 4.5 nm each in the center, assists in intracellular protein folding with the aid of GroES and adenosine triphosphate (ATP). Here, we investigated the possibility that GroEL can also encapsulate metal nanoparticles (NPs) up to ∼5 nm in diameter into the cavities with the aid of GroES and ATP. The slow ATP-hydrolyzing GroEL\u0000<sup>D52A/D398A</sup>\u0000 mutant, which forms extremely stable complexes with GroES (half-time of ∼6 days), made it possible to analyze GroEL/GroES complexes containing metal NPs. Scanning transmission electron microscopy–energy-dispersive X-ray spectroscopy analysis proved distinctly that FePt NPs and Au NPs were encapsulated in the GroEL/GroES complexes. Dynamic light scattering measurements showed that the NPs in the GroEL/GroES complex were able to maintain their dispersibility in solution. We previously described that the incubation of GroEL and GroES in the presence of ATP·BeFx and adenosine diphosphate·BeFx resulted in the formation of symmetric football-shaped and asymmetric bullet-shaped complexes, respectively. Based on this knowledge, we successfully constructed the football-shaped complex in which two compartments were occupied by Pt or Au NPs (first compartment) and FePt NPs (second compartment). This study showed that metal NPs were sequentially encapsulated according to the GroEL reaction in a step-by-step manner. In light of these results, chaperonin can be used as a tool for handling nanomaterials.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"289-296"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa064","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38683554","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}
引用次数: 1
Phase imaging dislocations using diffracted beam interferometry 用衍射束干涉法对位错进行相位成像
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa066
Rodney Herring
{"title":"Phase imaging dislocations using diffracted beam interferometry","authors":"Rodney Herring","doi":"10.1093/jmicro/dfaa066","DOIUrl":"10.1093/jmicro/dfaa066","url":null,"abstract":"A phase imaging method that measures the phase shift existing at a dislocation's core is described. The method uses the interference of two symmetrically diffracted beams on the optic axis by means of an electron biprism. Each diffracted beam carries half the phase of the dislocation core. When combined, the entire phase shift of the dislocation core is obtained.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"297-301"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa066","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38329842","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}
引用次数: 3
TEM observation of compacted DNA of Synechococcus elongatus PCC 7942 using DRAQ5 labeling with DAB photooxidation and osmium black DAB光氧化和锇黑DRAQ5标记细长聚球藻PCC 7942 DNA的透射电镜观察
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa058
Ilika Ghosh;Kimie Atsuzawa;Aoi Arai;Ryuzo Ohmukai;Yasuko Kaneko
{"title":"TEM observation of compacted DNA of Synechococcus elongatus PCC 7942 using DRAQ5 labeling with DAB photooxidation and osmium black","authors":"Ilika Ghosh;Kimie Atsuzawa;Aoi Arai;Ryuzo Ohmukai;Yasuko Kaneko","doi":"10.1093/jmicro/dfaa058","DOIUrl":"10.1093/jmicro/dfaa058","url":null,"abstract":"To visualize the fine structure of compacted DNA of Synechococcus elongatus PCC 7942, which appears at a specific time in the regular light/dark cycle prior to cell division, ChromEM with some modifications was applied. After staining DNA with DRAQ5, the cells were fixed and irradiated by red laser in the presence of 3,3ʹ-diaminobenzidine and subsequently fixed with OsO\u0000<inf>4</inf>\u0000. A system with He–Ne laser (633 nm) was set up for efficient irradiation of the bacterial cells in aqueous solution. The compacted DNA was visualized by transmission electron microscopy, in ultrathin sections as electron dense staining by osmium black.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"316-320"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa058","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38427660","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}
引用次数: 1
Ultrastructural evidence for an unusual mode of ciliogenesis in mouse multiciliated epithelia 小鼠多纤毛上皮纤毛发生异常模式的超微结构证据
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa074
Keishi Narita;Sen Takeda
{"title":"Ultrastructural evidence for an unusual mode of ciliogenesis in mouse multiciliated epithelia","authors":"Keishi Narita;Sen Takeda","doi":"10.1093/jmicro/dfaa074","DOIUrl":"10.1093/jmicro/dfaa074","url":null,"abstract":"Multiciliogenesis is a cascading process for generating hundreds of motile cilia in single cells. In vertebrates, this process has been investigated in the ependyma of brain ventricles and the ciliated epithelia of the airway and oviduct. Although the early steps to amplify centrioles have been characterized in molecular detail, subsequent steps to establish multicilia have been relatively overlooked. Here, we focused on unusual cilia-related structures previously observed in wild-type mouse ependyma using transmission electron microscopy and analyzed their ultrastructural features and the frequency of their occurrence. In the ependyma, \u0000<tex>$sim$</tex>\u00005% of cilia existed as bundles; while the majority of the bundles were paired, bundles of more than three cilia were also found. Furthermore, apical protrusions harboring multiple sets of axonemes were occasionally observed (0–2 per section), suggesting an unusual mode of ciliogenesis. In trachea and oviduct epithelia, ciliary bundles were absent, but protrusions containing multiple axonemes were observed. At the base of such protrusions, certain axonemes were completely enwrapped by membranes, whereas others remained incompletely enwrapped. These data suggested that the late steps of multiciliogenesis might include a unique process underlying the development of cilia, which is distinct from the ciliogenesis of primary cilia.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"308-315"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa074","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38660318","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}
引用次数: 2
Evaluation of depth of dislocation visibility in SEM electron channeling contrast imaging in Ti-6Al-4V alloy using serial sectioning method 用连续切片法评价Ti-6Al-4V合金SEM电子沟道对比成像中位错可见度的深度
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa060
Shigeto Yamasaki;Misaki Deguchi;Masatoshi Mitsuhara;Hideharu Nakashima;Yutaro Ota;Keiji Kubushiro
{"title":"Evaluation of depth of dislocation visibility in SEM electron channeling contrast imaging in Ti-6Al-4V alloy using serial sectioning method","authors":"Shigeto Yamasaki;Misaki Deguchi;Masatoshi Mitsuhara;Hideharu Nakashima;Yutaro Ota;Keiji Kubushiro","doi":"10.1093/jmicro/dfaa060","DOIUrl":"10.1093/jmicro/dfaa060","url":null,"abstract":"In this study, we conducted a quantitative evaluation of dislocation density by scanning electron microscopy electron channeling contrast imaging for α grains of a Ti-6Al-4V alloy deformed at room temperature. The depth of visibility of dislocations is experimentally measured as 140 to 160 nm by a serial sectioning observation. This result is compared with the theoretical value and applied to evaluate dislocation density. These factors confirm that the theoretically calculated value of the depth of visibility, at 5 to 6 times the extinction distance, is valid for the hexagonal close-packed Ti alloy.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"265-277"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa060","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38443091","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}
引用次数: 0
Photobleaching reduction in modulated super-resolution microscopy 调制超分辨率显微镜中的光漂白减少
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa062
Jafar H Ghithan;Jennifer M Noel;Thomas J Roussel;Maureen A McCall;Bruce W Alphenaar;Sergio B Mendes
{"title":"Photobleaching reduction in modulated super-resolution microscopy","authors":"Jafar H Ghithan;Jennifer M Noel;Thomas J Roussel;Maureen A McCall;Bruce W Alphenaar;Sergio B Mendes","doi":"10.1093/jmicro/dfaa062","DOIUrl":"10.1093/jmicro/dfaa062","url":null,"abstract":"Important breakthroughs in far-field imaging techniques have been made since the first demonstrations of stimulated emission depletion (STED) microscopy. To date, the most straightforward and widespread deployment of STED microscopy has used continuous wave (CW) laser beams for both the excitation and depletion of fluorescence emission. A major drawback of the CW STED imaging technique has been photobleaching effects due to the high optical power needed in the depletion beam to reach sub-diffraction resolution. To overcome this hurdle, we have applied a synchronous detection approach based on modulating the excitation laser beam, while keeping the depletion beam at CW operation, and frequency filtering the collected signal with a lock-in amplifier to record solely the super-resolved fluorescence emission. We demonstrate here that such approach allows an important reduction in the optical power of both laser beams that leads to measurable decreases in photobleaching effects in STED microscopy. We report super-resolution images with relatively low powers for both the excitation and depletion beams. In addition, typical unwanted scattering effects and background signal generated from the depletion beam, which invariably arises from mismatches in refractive index in the material composing the sample, are largely reduced by using the modulated STED approach. The capability of acquiring super-resolution images with relatively low power is quite relevant for studying a variety of samples, but particularly important for biological species as exemplified in this work.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"278-288"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa062","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38496307","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}
引用次数: 1
Denoising of series electron holograms using tensor decomposition 用张量分解对串联电子全息图进行去噪
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa057
Yuki Nomura;Kazuo Yamamoto;Satoshi Anada;Tsukasa Hirayama;Emiko Igaki;Koh Saitoh
{"title":"Denoising of series electron holograms using tensor decomposition","authors":"Yuki Nomura;Kazuo Yamamoto;Satoshi Anada;Tsukasa Hirayama;Emiko Igaki;Koh Saitoh","doi":"10.1093/jmicro/dfaa057","DOIUrl":"10.1093/jmicro/dfaa057","url":null,"abstract":"In this study, a noise-reduction technique for series low-dose electron holograms using tensor decomposition is demonstrated through simulation. We treated an entire dataset of the series holograms with Poisson noise as a third-order tensor, which is a stack of 2D holograms. The third-order tensor, which is decomposed into a core tensor and three factor matrices, is approximated as a lower-rank tensor using only noise-free principal components. This technique is applied to simulated holograms by assuming a p-n junction in a semiconductor sample. The peak signal-to-noise ratios of the holograms and the reconstructed phase maps have been improved significantly using tensor decomposition. Moreover, the proposed method was applied to a more practical situation of time-resolved in situ electron holography by considering a nonuniform fringe contrast and fringe drift relative to the sample. The accuracy and precision of the reconstructed phase maps were quantitatively evaluated to demonstrate its effectiveness for in situ experiments and low-dose experiments on beam-sensitive materials.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"255-264"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa057","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38396011","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}
引用次数: 4
Effect of nitrogen-containing bisphosphonates on osteoclasts and osteoclastogenesis: an ultrastructural study 含氮双磷酸盐对破骨细胞和破骨细胞生成的影响:超微结构研究
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa073
Yoshitoki Takagi;Satoshi Inoue;Kaoru Fujikawa;Miwako Matsuki-Fukushima;Mitsuori Mayahara;Kayo Matsuyama;Yasuo Endo;Masanori Nakamura
{"title":"Effect of nitrogen-containing bisphosphonates on osteoclasts and osteoclastogenesis: an ultrastructural study","authors":"Yoshitoki Takagi;Satoshi Inoue;Kaoru Fujikawa;Miwako Matsuki-Fukushima;Mitsuori Mayahara;Kayo Matsuyama;Yasuo Endo;Masanori Nakamura","doi":"10.1093/jmicro/dfaa073","DOIUrl":"10.1093/jmicro/dfaa073","url":null,"abstract":"We have previously indicated that a single injection of alendronate, one of the nitrogen-containing bisphosphonates (NBPs), affects murine hematopoietic processes, such as the shift of erythropoiesis from bone marrow (BM) to spleen, disappearance of BM-resident macrophages, the increase of granulopoiesis in BM and an increase in the number of osteoclasts. NBPs induce apoptosis and the formation of giant osteoclasts in vitro and/or in patients undergoing long-term NBP treatment. Therefore, the time-kinetic effect of NBPs on osteoclasts needs to be clarified. In this study, we examined the effect of alendronate on mouse osteoclasts and osteoclastogenesis. One day after the treatment, osteoclasts lost the clear zone and ruffled borders, and the cell size decreased. After 2 days, the cytoplasm of osteoclasts became electron dense and the nuclei became pyknotic. Some of the cells had fragmented nuclei. After 4 days, osteoclasts had euchromatic nuclei attached to the bone surface. Osteoclasts had no clear zones or ruffled borders. After 7 days, osteoclasts formed giant osteoclasts via the fusion of multinuclear and mononuclear osteoclasts. These results indicate that NBPs affect osteoclasts and osteoclastogenesis via two different mechanisms.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"302-307"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa073","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38675525","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}
引用次数: 4
Performance of a silicon-on-insulator direct electron detector in a low-voltage transmission electron microscope 低压透射电子显微镜中绝缘体上硅直接电子探测器的性能
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-06-01 DOI: 10.1093/jmicro/dfaa072
Takafumi Ishida;Akira Shinozaki;Makoto Kuwahara;Toshinobu Miyoshi;Koh Saitoh;Yasuo Arai
{"title":"Performance of a silicon-on-insulator direct electron detector in a low-voltage transmission electron microscope","authors":"Takafumi Ishida;Akira Shinozaki;Makoto Kuwahara;Toshinobu Miyoshi;Koh Saitoh;Yasuo Arai","doi":"10.1093/jmicro/dfaa072","DOIUrl":"10.1093/jmicro/dfaa072","url":null,"abstract":"The performance of a direct electron detector using silicon-on-insulator (SOI) technology in a low-voltage transmission electron microscope (LVTEM) is evaluated. The modulation transfer function and detective quantum efficiency of the detector are measured under backside illumination. The SOI-type detector is demonstrated to have high sensitivity and high efficiency for the direct detection of low-energy electrons. The detector is thus considered suitable for low-dose imaging in an LVTEM.","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"70 1","pages":"321-325"},"PeriodicalIF":1.8,"publicationDate":"2021-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1093/jmicro/dfaa072","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38592753","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}
引用次数: 1
Introduction to EBSD: Current Issues in Data Presentation. EBSD简介:数据表示中的当前问题。
IF 1.8 4区 工程技术
Microscopy Pub Date : 2021-05-01 DOI: 10.2172/1870763
JosephTan Michael
{"title":"Introduction to EBSD: Current Issues in Data Presentation.","authors":"JosephTan Michael","doi":"10.2172/1870763","DOIUrl":"https://doi.org/10.2172/1870763","url":null,"abstract":"","PeriodicalId":18515,"journal":{"name":"Microscopy","volume":"1 1","pages":""},"PeriodicalIF":1.8,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88336999","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}
引用次数: 0
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