MicroscopyPub Date : 2022-06-11DOI: 10.1093/jmicro/dfac028
Kaname Yoshida, Y. Sasaki, A. Kuwabara, Y. Ikuhara
{"title":"Reliable Electrochemical Setup for in situ Observations with an Atmospheric SEM.","authors":"Kaname Yoshida, Y. Sasaki, A. Kuwabara, Y. Ikuhara","doi":"10.1093/jmicro/dfac028","DOIUrl":"https://doi.org/10.1093/jmicro/dfac028","url":null,"abstract":"A novel setup for the in situ observation of electrochemical reactions in liquids through atmospheric scanning electron microscopy is presented. The proposed liquid-phase electrochemical SEM system consists of a working electrode (WE) on an electrochemical chip (e-chip) and other two electrodes inserted into a liquid electrolyte; electrochemical reactions occurring at the WE are controlled precisely with an external potentiostat/galvanostat connected to the three electrodes. Copper deposition from a CuSO4 aqueous solution was conducted onto the WE, and simultaneous acquisition of nanoscale images and reliable electrochemical data was achieved with the proposed setup.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46450430","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}
MicroscopyPub Date : 2022-05-30DOI: 10.1093/jmicro/dfac027
Dr. Saddam Hussain Khan, Najmus Saher Shah, Rabia Nuzhat, A. Majid, Hani Alquhayz, Asifullah Khan
{"title":"Malaria Parasite Classification Framework using a Novel Channel Squeezed and Boosted CNN.","authors":"Dr. Saddam Hussain Khan, Najmus Saher Shah, Rabia Nuzhat, A. Majid, Hani Alquhayz, Asifullah Khan","doi":"10.1093/jmicro/dfac027","DOIUrl":"https://doi.org/10.1093/jmicro/dfac027","url":null,"abstract":"Malaria is a life-threatening infection that infects the red blood cells (RBCs) that gradually grows throughout the body. The plasmodium parasite is caused by a female anopheles mosquito bite and severely affects numerous individuals within the world every year. Therefore, early detection tests are required to predict infected parasitic cells. The proposed technique exploits deep convolutional neural network (CNN) learning capability to detect the thin-blood smear parasitic patients from healthy individuals. In this regard, the detection is accomplished using a novel STM-SB-RENet block-based CNN that employs the idea of split-transform-merge (STM) and channel Squeezing-Boosting (SB) in a modified fashion. In this connection, a new convolutional block-based STM is developed, which systematically implements region and edge operations to explore the parasitic malaria pattern related to region-homogeneity, structural obstruction, and boundary-defining features. Moreover, the diverse boosted feature maps are achieved by incorporating the new channel SB and Transfer Learning (TL) idea in each STM block at abstract, intermediate, and target levels to capture minor contrast and texture variation between parasitic and normal artifacts. The malaria input images to the proposed models are initially transformed using discrete wavelet transform to generate enhanced and reduced feature space. The proposed architectures are validated using hold-out cross-validation on the National Institute of Health Malaria dataset. The proposed methods outperform the train from scratch, and TL-based fine-tuned existing techniques. The considerable performance (accuracy: 97.98%, sensitivity: 0.988, F-score: 0.980, and AUC: 0.996) of STM-SB-RENet suggests that it can be utilized to screen parasitic malaria patients.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42209666","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":"A Functional Platform for Remote use of Electron Microscopes Using Web Conferencing Systems.","authors":"Makoto Sugiura-Nakazato, Hiroshi Takase, Takeru Nakazato","doi":"10.1093/jmicro/dfac026","DOIUrl":"https://doi.org/10.1093/jmicro/dfac026","url":null,"abstract":"In this study, we propose a useful system for remote sharing of transmission electron microscope (TEM) images by connecting three computers: a computer connected to a TEM, a computer distributing images, and a computer receiving images. Then, we confirmed the performance of three web conferencing systems, Microsoft Teams, Zoom, and Google Meet, to evaluate the usefulness of their remote use based on the clarity of images, smoothness of movement, and time lag in images on each computer in the system. A screen image can be captured using the following two methods: a virtual camera of a video distribution software that provided a good reaction speed to transfer images and the screen sharing by conference system software that could share high-quality images.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44578214","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}
MicroscopyPub Date : 2022-05-10DOI: 10.1093/jmicro/dfac024
K. Johkura, N. Usuda, Yoshihiro Tanaka, Motoaki Fukasawa, K. Murata, T. Noda, N. Ohno
{"title":"Whole-cell observation of ZIO-stained Golgi apparatus in rat hepatocytes with serial block-face scanning electron microscope, SBF-SEM","authors":"K. Johkura, N. Usuda, Yoshihiro Tanaka, Motoaki Fukasawa, K. Murata, T. Noda, N. Ohno","doi":"10.1093/jmicro/dfac024","DOIUrl":"https://doi.org/10.1093/jmicro/dfac024","url":null,"abstract":"Abstract The Golgi apparatus, which plays a role in various biosynthetic pathways, is usually identified in electron microscopy by the morphological criteria of lamellae. A 3-dimensional analyses with serial block-face scanning electron microscope (SBF-SEM), a volume-SEM proficient in obtaining large volumes of data at the whole-cell level, could be a promising technique for understanding the precise distribution and complex ultrastructure of Golgi apparatus, although optimal methods for such analyses remain unclear since the observation can be hampered with sample charging and low image contrast, and manual segmentation often requires significant manpower. The present study attempted the whole-cell observation and semi-automatic classification and segmentation of the Golgi apparatus in rat hepatocytes for the first time by SBF-SEM via ZIO staining, a classical osmium impregnation. The staining electron-densely visualized individual Golgi lamellae, and their ultrastructure could stably be observed without any noticeable charging. The simple thresholding of the serial images enabled the efficient reconstruction of the labeled Golgi apparatus, which revealed plural Golgi apparatus in one hepatocyte. The combination of the heavy metal-based histochemistry of zinc, iodine and osmium (ZIO) staining and SBF-SEM was useful in the 3-dimensional observation of the Golgi apparatus at the whole-cell level because of two technical advantages: (i) visualization of the Golgi apparatus without any heavy metal staining and efficient acquisition of the block-face images without additional conductive staining or any devices for eliminating charging; (ii) easy identification of the staining and hassle-free, semi-automatic classification and segmentation by simple thresholding of the images. This novel approach could elucidate the topographic characteristics of the Golgi apparatus in hepatocytes.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42447308","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}
MicroscopyPub Date : 2022-05-04DOI: 10.1093/jmicro/dfac023
Y. Sasaki, Ayako Mizushima, Y. Mita, Kaname Yoshida, A. Kuwabara, Y. Ikuhara
{"title":"Design and Fabrication of an Electrochemical Chip for Liquid-Phase Transmission Electron Microscopy.","authors":"Y. Sasaki, Ayako Mizushima, Y. Mita, Kaname Yoshida, A. Kuwabara, Y. Ikuhara","doi":"10.1093/jmicro/dfac023","DOIUrl":"https://doi.org/10.1093/jmicro/dfac023","url":null,"abstract":"Liquid-phase transmission electron microscopy (LP-TEM) can be used with an electrochemical chip (e-chip) to observe electrochemical reactions in a liquid in situ. The design of electrodes on an e-chip fabricated using microelectromechanical system (MEMS) technology cannot be easily changed. Here, we report a newly designed e-chip and its fabrication process. Electrodes with a desired shape were fabricated with various metals via an additional step of vacuum deposition onto our e-chip with a shadow mask. For precise control of the electrochemical reactions in LP-TEM, optimization of the electrode shape and material is critical.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45071189","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}
MicroscopyPub Date : 2022-04-23DOI: 10.1093/jmicro/dfac021
T. Yamazaki, Hiromasa Niinomi, Y. Kimura
{"title":"Feasibility of control of particle assembly by dielectrophoresis in liquid-cell transmission electron microscopy","authors":"T. Yamazaki, Hiromasa Niinomi, Y. Kimura","doi":"10.1093/jmicro/dfac021","DOIUrl":"https://doi.org/10.1093/jmicro/dfac021","url":null,"abstract":"Abstract Liquid-cell transmission electron microscopy (LC-TEM) is a useful technique for observing phenomena in liquid samples with spatial and temporal resolutions similar to those of conventional transmission electron microscopy (TEM). This method is therefore expected to permit the visualization of phenomena previously inaccessible to conventional optical microscopy. However, dynamic processes such as nucleation are difficult to observe by this method because of difficulties in controlling the condition of the sample liquid in the observation area. To approach this problem, we focused on dielectrophoresis, in which electrodes are used to assemble particles, and we investigated the phenomena that occurred when an alternating-current signal was applied to an electrode in an existing liquid cell by using a phase-contrast optical microscope (PCM) and TEM. In PCM, we observed that colloidal particles in a solution were attracted to the electrodes to form assemblies, that the particles aligned along the electric field to form pearl chains and that the pearl chains accumulated to form colloidal crystals. However, these phenomena were not observed in the TEM study because of differences in the design of the relevant holders. The results of our study imply that the particle assembly by using dielectrophoretic forces in LC-TEM should be possible, but further studies, including electric device development, will be required to realize this in practice.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44674900","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}
MicroscopyPub Date : 2022-04-21DOI: 10.1093/jmicro/dfac020
Yuma Ozaki, T. Aimi, N. Shimomura
{"title":"Detection of Autophagy-Related Structures in Fruiting Bodies of Edible Mushroom, Pleurotus ostreatus.","authors":"Yuma Ozaki, T. Aimi, N. Shimomura","doi":"10.1093/jmicro/dfac020","DOIUrl":"https://doi.org/10.1093/jmicro/dfac020","url":null,"abstract":"Autophagy is involved in various fungal morphogenetic processes. However, there are limited reports regarding the role of autophagy in mushroom fruiting body formation. The purpose of this study was to reveal the autophagy-related structures in mushroom-forming fungi. The edible mushroom Pleurotus ostreatus was used in this study. Transmission electron microscopy revealed double-membrane bounded structures containing cytoplasmic components in the fruiting bodies of this fungus. Some of these double-membrane structures were observed to interact with the vacuoles. Additionally, curved flat cisternae of various lengths were detected in the cytoplasm. The shape, size, and thickness of the limiting membrane of the double-membrane structures and the flat cisternae corresponded well with those of the autophagosomes and the isolation membranes, respectively. Regarding autophagosome formation, a membrane-bound specific zone was detected near the isolation membrane, which appeared to expand along the novel membrane. This is the first detailed report showing autophagy-related structures in P. ostreatus and provides a possible model for autophagosome formation in these filamentous fungi. Mini-abstract Autophagy is involved in fungal morphogenetic processes. The fruiting bodies of edible mushroom Pleurotus ostreatus was observed under a TEM. The present study showed autophagy-related structures in this fungus and provides a possible model for autophagosome formation in filamentous fungi.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44591942","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}
MicroscopyPub Date : 2022-04-08DOI: 10.1093/jmicro/dfac017
Tomoko Kamasaki, Ryota Uehara, Yasuyuki Fujita
{"title":"Ultrastructural characteristics of finger-like membrane protrusions in cell competition","authors":"Tomoko Kamasaki, Ryota Uehara, Yasuyuki Fujita","doi":"10.1093/jmicro/dfac017","DOIUrl":"https://doi.org/10.1093/jmicro/dfac017","url":null,"abstract":"Abstract A small number of oncogenic mutated cells sporadically arise within the epithelial monolayer. Newly emerging Ras- or Src-transformed epithelial cells are often apically eliminated during competitive interactions between normal and transformed cells. Our recent electron microscopy (EM) analyses revealed that characteristic finger-like membrane protrusions are formed at the interface between normal and RasV12-transformed cells via the cdc42–formin-binding protein 17 (FBP17) pathway, potentially playing a positive role in intercellular recognition during apical extrusion. However, the spatial distribution and ultrastructural characteristics of finger-like protrusions remain unknown. In this study, we performed both X–Y and X–Z EM analyses of finger-like protrusions during the apical extrusion of RasV12-transformed cells. Quantification of the distribution and widths of the protrusions showed comparable results between the X–Y and X–Z sections. Finger-like protrusions were observed throughout the cell boundary between normal and RasV12 cells, except for apicalmost tight junctions. In addition, a non-cell-autonomous reduction in protrusion widths was observed between RasV12 cells and surrounding normal cells under the mix culture condition. In the finger-like protrusions, intercellular adhesions via thin electron-dense plaques were observed, implying that immature and transient forms of desmosomes, adherens junctions or unknown weak adhesions were distributed. Interestingly, unlike RasV12-transformed cells, Src-transformed cells form fewer evident protrusions, and FBP17 in Src cells is dispensable for apical extrusion. Collectively, these results suggest that the dynamic reorganization of intercellular adhesions via finger-like protrusions may positively control cell competition between normal and RasV12-transformed cells. Furthermore, our data indicate a cell context–dependent diversity in the modes of apical extrusion.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46296015","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}
MicroscopyPub Date : 2022-03-29DOI: 10.1093/jmicro/dfac016
Yuji Kanazawa, M. Nagano, S. Koinuma, S. Sugiyo, Y. Shigeyoshi
{"title":"Effects of aging on basement membrane of tibialis anterior muscle during recovery following muscle injury in rats","authors":"Yuji Kanazawa, M. Nagano, S. Koinuma, S. Sugiyo, Y. Shigeyoshi","doi":"10.1093/jmicro/dfac016","DOIUrl":"https://doi.org/10.1093/jmicro/dfac016","url":null,"abstract":"Abstract We investigated the effect of aging on the basement membrane (BM) during postinjury muscle recovery. Using a rat model, we found that aging delayed muscle fiber and BM recovery. In addition, expression of BM-related factors peaked 7 days after muscle injury among both young and older rats. Peak expression of collagen IV synthetic factors decreased with age, whereas expression of the degradative factor was unaffected by age. These results suggest that age-related delays in postinjury muscle fiber and BM recovery may be related to the suppression of collagen IV synthetic factors.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60920873","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}
MicroscopyPub Date : 2022-03-11DOI: 10.1093/jmicro/dfac012
Xiaoguang Li, K. Mitsuishi, M. Takeguchi
{"title":"Effect of Amorphous Carbon Coating on the Performance of Liquid Phase Transmission Electron Microscopy (LP-TEM) and the Dynamics of Enclosed Pt Nano-Colloids.","authors":"Xiaoguang Li, K. Mitsuishi, M. Takeguchi","doi":"10.1093/jmicro/dfac012","DOIUrl":"https://doi.org/10.1093/jmicro/dfac012","url":null,"abstract":"Ultra-thin silicon nitride (SiN) membranes are critical in microfabrication-based liquid cells (LCs) for transmission electron microscopy. This study used a homemade LC with a 50-nm SiN membrane to study the dynamics of 2.58-nm platinum (Pt) nanoparticles (NPs) in approximately 200-nm deep water. When a strong beam with electron flux ranging from 2.5 × 103 to 1.4 ×106 e-/(nm2·s) was applied to resolve the NPs, the beam caused NP aggregation and even drilled a hole on the top membrane. The hole drilling was prevented by coating a 1-4-nm-thick amorphous carbon layer on both sides of the membrane. The NP aggregation rate also decreased with increasing carbon thickness. After overcoming the aforementioned issues, lattice fringes of the Pt NPs were visible when the NPs were attached to the membrane of the 4-nm-carbon-coated LC containing a thin liquid layer. The effects of the electron beam and carbon on the LC and Pt NPs were investigated and discussed. This work provides a reference for LC-TEM research using strong electron beams.","PeriodicalId":48655,"journal":{"name":"Microscopy","volume":null,"pages":null},"PeriodicalIF":1.8,"publicationDate":"2022-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46772698","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}