Microscopy (Oxford, England)最新文献_第7页

A tool for live-cell confocal imaging of temperature-dependent organelle dynamics. 对温度依赖性细胞器动态进行活细胞共聚焦成像的工具。

Microscopy (Oxford, England) Pub Date : 2024-07-30 DOI: 10.1093/jmicro/dfad064

Keiko Midorikawa, Yutaka Kodama

引用次数: 0

Axonal regrowth under release of myelin-associated glycoprotein: chemotaxis by pioneer Schwann cells and Cajal's gigantic clubs. 髓鞘相关糖蛋白释放下的轴突再生：先锋雪旺细胞和卡哈尔巨型俱乐部的趋化作用。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad046

Kojun Torigoe

{"title":"Axonal regrowth under release of myelin-associated glycoprotein: chemotaxis by pioneer Schwann cells and Cajal's gigantic clubs.","authors":"Kojun Torigoe","doi":"10.1093/jmicro/dfad046","DOIUrl":"10.1093/jmicro/dfad046","url":null,"abstract":"Myelin-associated glycoprotein (MAG), released from pre-degenerated distal nerves following axotomy, blocks the regrowth of sprouts and naked axons. Ensheathed axons, however, continue to elongate and reach MAG-releasing distal nerves. To determine the regenerative mechanism of ensheathed axons without navigators of axonal growth cones by the film model method, we inserted a MAG-releasing distal nerve segment treated with liquid nitrogen (N2DS) between the two films, facing the proximal end of the common peroneal nerves in mice transected 4 days earlier for axons to become ensheathed. On the third postoperative day (Day 3), axon fascicles, subjected to silver staining, extended toward N2DS but with few branches, forming terminal swellings called Cajal's gigantic clubs (CGCs) that are filled with axonal growth cones. Filter paper wetted with either 250 pg/ml MAG or N2DS showed the same configurations when inserted between the two films. This effect was lost following anti-MAG treatment; fascicles strayed near the parent nerve with numerous branches, formed a net of axons and tapered toward thin tips at their ends, just like controls without N2DS. Schwann cell bundles on Day 3 detected with anti-S100, formed sheaths of CGCs at their ends and connected to pioneer Schwann cells (pSCs). To analyze the physiology of Schwann cells, independent of axons, the parent nerve transected 4 days prior was crushed. On Day 2, with pSCs ahead, Schwann cell bundles extended toward N2DS. On Day 4, main bundles regressed, leaving pSCs motionless. Thus, MAG is a candidate chemoattractant for both pSCs and CGCs.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"251-261"},"PeriodicalIF":0.0,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41156766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Three-dimensional analysis of the intracellular architecture by scanning electron microscopy. 通过扫描电子显微镜对细胞内结构进行三维分析。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad050

Daisuke Koga, Satoshi Kusumi, Hirokazu Yagi, Koichi Kato

{"title":"Three-dimensional analysis of the intracellular architecture by scanning electron microscopy.","authors":"Daisuke Koga, Satoshi Kusumi, Hirokazu Yagi, Koichi Kato","doi":"10.1093/jmicro/dfad050","DOIUrl":"10.1093/jmicro/dfad050","url":null,"abstract":"The two-dimensional observation of ultrathin sections from resin-embedded specimens provides an insufficient understanding of the three-dimensional (3D) morphological information of membranous organelles. The osmium maceration method, developed by Professor Tanaka's group >40 years ago, is the only technique that allows direct observation of the 3D ultrastructure of membrane systems using scanning electron microscopy (SEM), without the need for any reconstruction process. With this method, the soluble cytoplasmic proteins are removed from the freeze-cracked surface of cells while preserving the integrity of membranous organelles, achieved by immersing tissues in a diluted osmium solution for several days. By employing the maceration method, researchers using SEM have revealed the 3D ultrastructure of organelles such as the Golgi apparatus, mitochondria and endoplasmic reticulum in various cell types. Recently, we have developed new SEM techniques based on the maceration method to explore further possibilities of this method. These include: (i) a rapid osmium maceration method that reduces the reaction duration of the procedure, (ii) a combination method that combines agarose embedding with osmium maceration to elucidate the 3D ultrastructure of organelles in free and cultured cells and (iii) a correlative immunofluorescence and SEM technique that combines cryosectioning with the osmium maceration method, enabling the correlation of the immunocytochemical localization of molecules with the 3D ultrastructure of organelles. In this paper, we review the novel osmium maceration methods described earlier and discuss their potential and future directions in the field of biology and biomedical research.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"215-225"},"PeriodicalIF":0.0,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71489684","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Toward quantitative super-resolution microscopy: molecular maps with statistical guarantees. 走向定量超分辨率显微镜:具有统计保证的分子图谱。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad053

Katharina Proksch, Frank Werner, Jan Keller-Findeisen, Haisen Ta, Axel Munk

{"title":"Toward quantitative super-resolution microscopy: molecular maps with statistical guarantees.","authors":"Katharina Proksch, Frank Werner, Jan Keller-Findeisen, Haisen Ta, Axel Munk","doi":"10.1093/jmicro/dfad053","DOIUrl":"10.1093/jmicro/dfad053","url":null,"abstract":"Quantifying the number of molecules from fluorescence microscopy measurements is an important topic in cell biology and medical research. In this work, we present a consecutive algorithm for super-resolution (stimulated emission depletion (STED)) scanning microscopy that provides molecule counts in automatically generated image segments and offers statistical guarantees in form of asymptotic confidence intervals. To this end, we first apply a multiscale scanning procedure on STED microscopy measurements of the sample to obtain a system of significant regions, each of which contains at least one molecule with prescribed uniform probability. This system of regions will typically be highly redundant and consists of rectangular building blocks. To choose an informative but non-redundant subset of more naturally shaped regions, we hybridize our system with the result of a generic segmentation algorithm. The diameter of the segments can be of the order of the resolution of the microscope. Using multiple photon coincidence measurements of the same sample in confocal mode, we are then able to estimate the brightness and number of molecules and give uniform confidence intervals on the molecule counts for each previously constructed segment. In other words, we establish a so-called molecular map with uniform error control. The performance of the algorithm is investigated on simulated and real data.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"287-300"},"PeriodicalIF":0.0,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138178186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optical microscopic imaging, manipulation, and analysis methods for morphogenesis research. 用于形态发生研究的光学显微成像、操作和分析方法。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad059

Takanobu A Katoh, Yohsuke T Fukai, Tomoki Ishibashi

{"title":"Optical microscopic imaging, manipulation, and analysis methods for morphogenesis research.","authors":"Takanobu A Katoh, Yohsuke T Fukai, Tomoki Ishibashi","doi":"10.1093/jmicro/dfad059","DOIUrl":"10.1093/jmicro/dfad059","url":null,"abstract":"Morphogenesis is a developmental process of organisms being shaped through complex and cooperative cellular movements. To understand the interplay between genetic programs and the resulting multicellular morphogenesis, it is essential to characterize the morphologies and dynamics at the single-cell level and to understand how physical forces serve as both signaling components and driving forces of tissue deformations. In recent years, advances in microscopy techniques have led to improvements in imaging speed, resolution and depth. Concurrently, the development of various software packages has supported large-scale, analyses of challenging images at the single-cell resolution. While these tools have enhanced our ability to examine dynamics of cells and mechanical processes during morphogenesis, their effective integration requires specialized expertise. With this background, this review provides a practical overview of those techniques. First, we introduce microscopic techniques for multicellular imaging and image analysis software tools with a focus on cell segmentation and tracking. Second, we provide an overview of cutting-edge techniques for mechanical manipulation of cells and tissues. Finally, we introduce recent findings on morphogenetic mechanisms and mechanosensations that have been achieved by effectively combining microscopy, image analysis tools and mechanical manipulation techniques.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"226-242"},"PeriodicalIF":0.0,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11154147/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138812986","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Phase retrieval of electron rocking curves using total variation and total squared variation regularizations. 使用全变差和全平方变差正则化的电子摇摆曲线的相位反演。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad048

Akihiro Shichi, Hiroyuki Ishizuka, Koh Saitoh

引用次数: 0

Memory-efficient semantic segmentation of large microscopy images using graph-based neural networks. 使用基于图的神经网络对大型显微镜图像进行记忆高效的语义分割。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad049

Atishay Jain, David H Laidlaw, Peter Bajcsy, Ritambhara Singh

{"title":"Memory-efficient semantic segmentation of large microscopy images using graph-based neural networks.","authors":"Atishay Jain, David H Laidlaw, Peter Bajcsy, Ritambhara Singh","doi":"10.1093/jmicro/dfad049","DOIUrl":"10.1093/jmicro/dfad049","url":null,"abstract":"We present a graph neural network (GNN)-based framework applied to large-scale microscopy image segmentation tasks. While deep learning models, like convolutional neural networks (CNNs), have become common for automating image segmentation tasks, they are limited by the image size that can fit in the memory of computational hardware. In a GNN framework, large-scale images are converted into graphs using superpixels (regions of pixels with similar color/intensity values), allowing us to input information from the entire image into the model. By converting images with hundreds of millions of pixels to graphs with thousands of nodes, we can segment large images using memory-limited computational resources. We compare the performance of GNN- and CNN-based segmentation in terms of accuracy, training time and required graphics processing unit memory. Based on our experiments with microscopy images of biological cells and cell colonies, GNN-based segmentation used one to three orders-of-magnitude fewer computational resources with only a change in accuracy of ‒2 % to +0.3 %. Furthermore, errors due to superpixel generation can be reduced by either using better superpixel generation algorithms or increasing the number of superpixels, thereby allowing for improvement in the GNN framework's accuracy. This trade-off between accuracy and computational cost over CNN models makes the GNN framework attractive for many large-scale microscopy image segmentation tasks in biology.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"275-286"},"PeriodicalIF":0.0,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49685858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Contrast mechanism at landing energy near 0 eV in super low-energy scanning electron microscopy. 超低能量扫描电子显微镜在着陆能量接近 0 eV 时的对比机制。

Microscopy (Oxford, England) Pub Date : 2024-06-06 DOI: 10.1093/jmicro/dfad042

Tomohiro Aoyama, Šárka Mikmeková, Kazuhiro Kumagai

{"title":"Contrast mechanism at landing energy near 0 eV in super low-energy scanning electron microscopy.","authors":"Tomohiro Aoyama, Šárka Mikmeková, Kazuhiro Kumagai","doi":"10.1093/jmicro/dfad042","DOIUrl":"10.1093/jmicro/dfad042","url":null,"abstract":"In recent years, the technique of scanning electron microscopy (SEM) observation with low landing energy of a few keV or less has become common. We have especially focused on the drastic contrast change at near 0 eV. Using a patterned sample consisting of Si, Ni and Pt, threshold energies where the total reflection of incident electrons occurs were investigated by SEM at near 0 eV. In both the cases of in-situ and ex-situ sample cleaning, drastic changes in the brightness of each material were observed at near 0 eV, with threshold energies in the order Si < Ni < Pt. This order agreed with the order of the literature values of the work functions and the surface potentials measured by Kelvin force probe microscopy. This result suggests that the difference of the threshold energy is caused by the difference in surface potential due to the work function difference of each material. Although the order of the threshold energies also agreed with those of work functions reported in the literature, the work functions of air-exposed surfaces should be rather considered as 'modified work functions', since they could be significantly altered by the adsorbates, etc. Nevertheless, the difference of the threshold energy for each material was observed with commercial SEM at landing energy near 0 eV, which opens a new possibility to distinguish materials, although the difference should be rather recognized as 'fingerprints', since surface potentials are sensitive to conditions of surface treatments and atmospheric exposure.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"243-250"},"PeriodicalIF":0.0,"publicationDate":"2024-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10067988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Simultaneous secondary electron microscopy in the scanning transmission electron microscope with applications for in situ studies. 扫描透射电子显微镜中的同步二次电子显微镜，以及在原位研究中的应用。

Microscopy (Oxford, England) Pub Date : 2024-04-08 DOI: 10.1093/jmicro/dfae007

Mia L San Gabriel, Chenyue Qiu, Dian Yu, Toshie Yaguchi, Jane Y Howe

{"title":"Simultaneous secondary electron microscopy in the scanning transmission electron microscope with applications for in situ studies.","authors":"Mia L San Gabriel, Chenyue Qiu, Dian Yu, Toshie Yaguchi, Jane Y Howe","doi":"10.1093/jmicro/dfae007","DOIUrl":"10.1093/jmicro/dfae007","url":null,"abstract":"Scanning/transmission electron microscopy (STEM) is a powerful characterization tool for a wide range of materials. Over the years, STEMs have been extensively used for in situ studies of structural evolution and dynamic processes. A limited number of STEM instruments are equipped with a secondary electron (SE) detector in addition to the conventional transmitted electron detectors, i.e. the bright-field (BF) and annular dark-field (ADF) detectors. Such instruments are capable of simultaneous BF-STEM, ADF-STEM and SE-STEM imaging. These methods can reveal the 'bulk' information from BF and ADF signals and the surface information from SE signals for materials <200 nm thick. This review first summarizes the field of in situ STEM research, followed by the generation of SE signals, SE-STEM instrumentation and applications of SE-STEM analysis. Combining with various in situ heating, gas reaction and mechanical testing stages based on microelectromechanical systems (MEMS), we show that simultaneous SE-STEM imaging has found applications in studying the dynamics and transient phenomena of surface reconstructions, exsolution of catalysts, lunar and planetary materials and mechanical properties of 2D thin films. Finally, we provide an outlook on the potential advancements in SE-STEM from the perspective of sample-related factors, instrument-related factors and data acquisition and processing.","PeriodicalId":74193,"journal":{"name":"Microscopy (Oxford, England)","volume":" ","pages":"169-183"},"PeriodicalIF":0.0,"publicationDate":"2024-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139708760","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In situ TEM studies on hydrogen-related issues: hydrogen storage, hydrogen embrittlement, fuel cells and electrolysis. 氢相关问题的原位 TEM 研究：氢储存、氢脆化、燃料电池和电解。

Microscopy (Oxford, England) Pub Date : 2024-04-08 DOI: 10.1093/jmicro/dfad060

Junko Matsuda

引用次数: 0