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The LaserFIB: new application opportunities combining a high-performance FIB-SEM with femtosecond laser processing in an integrated second chamber LaserFIB:在集成的第二腔室中,将高性能FIB-SEM与飞秒激光处理相结合,提供了新的应用机会
Q3 Immunology and Microbiology Pub Date : 2020-10-26 DOI: 10.1186/s42649-020-00044-5
Ben Tordoff, Cheryl Hartfield, Andrew J. Holwell, Stephan Hiller, Marcus Kaestner, Stephen Kelly, Jaehan Lee, Sascha Müller, Fabian Perez-Willard, Tobias Volkenandt, Robin White, Thomas Rodgers

The development of the femtosecond laser (fs laser) with its ability to provide extremely rapid athermal ablation of materials has initiated a renaissance in materials science. Sample milling rates for the fs laser are orders of magnitude greater than that of traditional focused ion beam (FIB) sources currently used. In combination with minimal surface post-processing requirements, this technology is proving to be a game changer for materials research. The development of a femtosecond laser attached to a focused ion beam scanning electron microscope (LaserFIB) enables numerous new capabilities, including access to deeply buried structures as well as the production of extremely large trenches, cross sections, pillars and TEM H-bars, all while preserving microstructure and avoiding or reducing FIB polishing. Several high impact applications are now possible due to this technology in the fields of crystallography, electronics, mechanical engineering, battery research and materials sample preparation. This review article summarizes the current opportunities for this new technology focusing on the materials science megatrends of engineering materials, energy materials and electronics.

飞秒激光器(fs激光器)的发展及其提供极快的材料非热烧蚀的能力已经开始了材料科学的复兴。光纤激光器的样品铣削速率比目前使用的传统聚焦离子束(FIB)源的铣削速率大几个数量级。结合最小的表面后处理要求,这项技术被证明是材料研究的游戏规则改变者。将飞秒激光器连接到聚焦离子束扫描电子显微镜(LaserFIB)上,实现了许多新功能,包括进入深埋结构,以及生产极大的沟槽、横截面、柱和TEM h棒,同时保持微观结构,避免或减少FIB抛光。由于该技术在晶体学,电子学,机械工程,电池研究和材料样品制备领域的一些高影响应用现在成为可能。本文从工程材料、能源材料和电子材料的材料科学大趋势等方面综述了该新技术的发展前景。
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引用次数: 19
Noise2Atom: unsupervised denoising for scanning transmission electron microscopy images Noise2Atom:扫描透射电子显微镜图像的无监督去噪
Q3 Immunology and Microbiology Pub Date : 2020-10-20 DOI: 10.1186/s42649-020-00041-8
Feng Wang, Trond R. Henninen, Debora Keller, Rolf Erni

We propose an effective deep learning model to denoise scanning transmission electron microscopy (STEM) image series, named Noise2Atom, to map images from a source domain (mathcal {S}) to a target domain (mathcal {C}), where (mathcal {S}) is for our noisy experimental dataset, and (mathcal {C}) is for the desired clear atomic images. Noise2Atom uses two external networks to apply additional constraints from the domain knowledge. This model requires no signal prior, no noise model estimation, and no paired training images. The only assumption is that the inputs are acquired with identical experimental configurations. To evaluate the restoration performance of our model, as it is impossible to obtain ground truth for our experimental dataset, we propose consecutive structural similarity (CSS) for image quality assessment, based on the fact that the structures remain much the same as the previous frame(s) within small scan intervals. We demonstrate the superiority of our model by providing evaluation in terms of CSS and visual quality on different experimental datasets.

我们提出了一个有效的深度学习模型来对扫描透射电子显微镜(STEM)图像序列进行降噪,名为Noise2Atom,将图像从源域(mathcal {S})映射到目标域(mathcal {C}),其中(mathcal {S})用于我们的噪声实验数据集,(mathcal {C})用于所需的清晰原子图像。Noise2Atom使用两个外部网络来应用来自领域知识的附加约束。该模型不需要信号先验,不需要噪声模型估计,不需要成对训练图像。唯一的假设是输入是用相同的实验配置获得的。为了评估我们的模型的恢复性能,因为我们的实验数据集不可能获得地面真实值,我们提出了连续结构相似性(CSS)用于图像质量评估,基于这样一个事实,即结构在很小的扫描间隔内与前一帧保持大致相同。我们通过在不同的实验数据集上提供CSS和视觉质量方面的评估来证明我们模型的优越性。
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引用次数: 28
Method of Ga removal from a specimen on a microelectromechanical system-based chip for in-situ transmission electron microscopy 用于原位透射电子显微镜的基于微机电系统芯片的样品Ga去除方法
Q3 Immunology and Microbiology Pub Date : 2020-10-14 DOI: 10.1186/s42649-020-00043-6
Yena Kwon, Byeong-Seon An, Yeon-Ju Shin, Cheol-Woong Yang

In-situ transmission electron microscopy (TEM) holders that employ a chip-type specimen stage have been widely utilized in recent years. The specimen on the microelectromechanical system (MEMS)-based chip is commonly prepared by focused ion beam (FIB) milling and ex-situ lift-out (EXLO). However, the FIB-milled thin-foil specimens are inevitably contaminated with Ga+ ions. When these specimens are heated for real time observation, the Ga+ ions influence the reaction or aggregate in the protection layer. An effective method of removing the Ga residue by Ar+ ion milling within FIB system was explored in this study. However, the Ga residue remained in the thin-foil specimen that was extracted by EXLO from the trench after the conduct of Ar+ ion milling. To address this drawback, the thin-foil specimen was attached to an FIB lift-out grid, subjected to Ar+ ion milling, and subsequently transferred to an MEMS-based chip by EXLO. The removal of the Ga residue was confirmed by energy dispersive spectroscopy.

采用芯片式试样台的原位透射电子显微镜(TEM)支架近年来得到了广泛应用。基于微机电系统(MEMS)的芯片上的试样通常采用聚焦离子束(FIB)铣削和非原位提升(EXLO)法制备。然而,fib铣削薄片试样不可避免地受到Ga+离子的污染。当这些试样被加热进行实时观察时,Ga+离子会影响反应或聚集在保护层中。本研究探索了在FIB系统中利用Ar+离子铣削去除Ga残渣的有效方法。而氩离子铣削后用EXLO提取的薄片样品中仍有Ga残留。为了解决这一缺陷,薄片试样被附着在FIB吊出网格上,进行Ar+离子铣削,随后通过EXLO转移到基于mems的芯片上。通过能量色散谱分析证实了Ga残留的去除。
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引用次数: 1
Sublimable materials facilitate the TEM sample preparation of oil-soluble nanomaterials 可升华材料有利于油溶性纳米材料的TEM样品制备
Q3 Immunology and Microbiology Pub Date : 2020-09-29 DOI: 10.1186/s42649-020-00042-7
Yu-Hao Deng

Sample preparation is significantly important to the high-resolution transmission electron microscopy (HRTEM) characterization of nanomaterials. However, many general organic solvents can dissolve the necessary organic polymer support layer in TEM grid, which causes it difficult to obtain high-quality samples of oil-soluble nanomaterials. In this study, a new sample preparation method for oil-soluble nanomaterials has been developed by using the sublimable material as a transition layer. Experiments also show that there is no damage to TEM grids and high-quality HRTEM images can be obtained via this method. This approach paves the way to applicable HRTEM sample preparation of oil-soluble nanomaterials.

样品制备对纳米材料的高分辨率透射电镜(HRTEM)表征具有重要意义。然而,许多普通的有机溶剂可以溶解TEM网格中必要的有机聚合物支撑层,导致难以获得高质量的油溶性纳米材料样品。本研究开发了一种以可升华材料为过渡层制备油溶性纳米材料样品的新方法。实验结果表明,该方法对TEM网格无损伤,可获得高质量的HRTEM图像。该方法为应用HRTEM制备油溶性纳米材料的样品铺平了道路。
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引用次数: 1
Fine structure of the intercalated disc and cardiac junctions in the black widow spider Latrodectus mactans 黑寡妇蜘蛛间插盘和心脏连接处的精细结构
Q3 Immunology and Microbiology Pub Date : 2020-09-25 DOI: 10.1186/s42649-020-00040-9
Yan Sun, Seung-Min Lee, Bon-Jin Ku, Myung-Jin Moon

Arthropods have an open circulatory system with a simple tubular heart, so it has been estimated that the contractile pumping structure of the cardiac muscle will be less efficient than that of vertebrates. Nevertheless, certain arthropods are known to have far superior properties and characteristics than vertebrates, so we investigated the fine structural features of intercalated discs and cardiac junctions of cardiac muscle cells in the black widow spider Latrodectus mactans. Characteristically, the spider cardiac muscle has typical striated features and represents a functional syncytium that supports multiple connections to adjacent cells by intercalated discs. Histologically, the boundary lamina of each sarcolemma connects to the basement membrane to form an elastic sheath, and the extracellular matrix allows the cells to be anchored to other tissues. Since the intercalated disc is also part of sarcolemma, it contains gap junctions for depolarization and desmosomes that keep the fibers together during cardiac muscle contraction. Furthermore, fascia adherens and macula adherens (desmosomes) were also identified as cell junctions in both sarcolemma and intercalated discs. To enable the coordinated heartbeat of the cardiac muscle, the muscle fibers have neuronal innervations by multiple axons from the motor ganglion.

节肢动物有一个开放的循环系统和一个简单的管状心脏,因此据估计,心肌的收缩泵结构将比脊椎动物的效率低。然而,已知某些节肢动物具有比脊椎动物优越得多的特性和特征,因此我们研究了黑寡妇蜘蛛(Latrodectus mactans)心肌细胞间插盘和心脏连接处的精细结构特征。蜘蛛心肌具有典型的条纹特征,是一种功能性合胞体,通过插片支持与相邻细胞的多重连接。组织学上,每个肌膜的边界层与基底膜连接形成弹性鞘,细胞外基质允许细胞锚定在其他组织上。由于间插椎间盘也是肌膜的一部分,它包含用于去极化的间隙连接和在心肌收缩时使纤维保持在一起的桥粒。此外,筋膜粘附体和黄斑粘附体(桥粒)也被鉴定为肌膜和间插椎间盘的细胞连接。为了使心肌的协调心跳,肌肉纤维受到来自运动神经节的多个轴突的神经支配。
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引用次数: 2
Methods to evaluate the twin formation energy: comparative studies of the atomic simulations and in-situ TEM tensile tests 评价孪晶形成能的方法:原子模拟与原位TEM拉伸试验的对比研究
Q3 Immunology and Microbiology Pub Date : 2020-09-17 DOI: 10.1186/s42649-020-00039-2
Hong-Kyu Kim, Sung-Hoon Kim, Jae-Pyoung Ahn

Deformation twinning, one of the major deformation modes in a crystalline material, has typically been analyzed using generalized planar fault energy (GPFE) curves. Despite the significance of these curves in understanding the twin nucleation and its effect on the mechanical properties of crystals, their experimental validity is lacking. In this comparative study based on the first-principles calculation, molecular dynamics simulation, and quantitative in-situ tensile testing of Al nanowires inside a transmission electron microscopy system, we present both a theoretical and an experimental approach that enable the measurement of a part of the twin formation energy of the perfect Al crystal. The proposed experimental method is also regarded as an indirect but quantitative means for validating the GPFE theory.

变形孪晶是晶体材料的主要变形模式之一,通常采用广义平面断层能(GPFE)曲线进行分析。尽管这些曲线在理解孪核及其对晶体力学性能的影响方面具有重要意义,但它们的实验有效性不足。在这项基于第一性原理计算、分子动力学模拟和透射电子显微镜系统内铝纳米线的原位定量拉伸测试的比较研究中,我们提出了一种理论和实验方法,可以测量完美铝晶体的部分孪晶形成能量。所提出的实验方法也被认为是验证GPFE理论的间接但定量的手段。
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引用次数: 1
Microscopic research on the olfactory organ of the Far Eastern brook lamprey Lethenteron reissneri (Pisces, Petromyzontidae) 远东七鳃鳗嗅觉器官的显微研究(双鱼座,七鳃鳗科)
Q3 Immunology and Microbiology Pub Date : 2020-09-15 DOI: 10.1186/s42649-020-00038-3
Hyun-Tae Kim, Jong-Young Park

The olfactory anatomy and histology of Lethenteron reissneri were researched using a stereo microscope, a light microscope, and a scanning electron microscope. As in other lampreys, it shows same characters as follows: i) a single olfactory organ, ii) a single tubular nostril, iii) a single olfactory chamber with gourd-like form, iv) a nasal valve, v) a nasopharyngeal pouch, vi) a sensory epithelium (SE) of continuous distribution, vii) a supporting cells with numerous long cilia, viii) an accessory olfactory organ. However, the description of a pseudostratified columnar layer in the SE and Non SE is a first record, not reported in sea lamprey Petromyzon marinus. In particular, both 19 to 20 lamellae in number and olfactory receptor neuron’s quarter ciliary length of the knob diameter differ from those of P. marinus. From these results, it might be considered that the olfactory organ of L. reissneri shows well adaptive structure of a primitive fish to slow flowing water with gravel, pebbles, and sand and a hiding habit into sand bottom at daytime. The lamellar number and neuron’s ciliary length may be a meaningful taxonomic character for the class Petromyzonida.

采用立体显微镜、光学显微镜和扫描电镜对雷氏肉鸡的嗅觉解剖和组织学进行了研究。与其他七鳃鳗相同,七鳃鳗具有以下特征:1)单个嗅觉器官,2)单个管状鼻孔,3)单个葫芦状嗅室,4)鼻阀,5)鼻咽袋,6)连续分布的感觉上皮,7)有大量长纤毛的支撑细胞,8)副嗅觉器官。然而,在东南和非东南的假层状柱状层的描述是首次记录,在海洋七鳃鳗中未见报道。特别是19 ~ 20片叶的数量和旋钮直径嗅觉受体神经元的四分之一纤毛长度都与海苔不同。从这些结果可以认为,雷氏水蛭的嗅觉器官表现出一种原始鱼类对含砾石、卵石和沙子的缓慢流动的水的良好适应结构和白天躲在沙底的习性。片层数和神经元纤毛长度可能是岩裂虫纲的一个有意义的分类特征。
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引用次数: 1
Depth-dependent EBIC microscopy of radial-junction Si micropillar arrays 径向结硅微柱阵列的深度依赖EBIC显微镜
Q3 Immunology and Microbiology Pub Date : 2020-09-03 DOI: 10.1186/s42649-020-00037-4
Kaden M. Powell, Heayoung P. Yoon

Recent advances in fabrication have enabled radial-junction architectures for cost-effective and high-performance optoelectronic devices. Unlike a planar PN junction, a radial-junction geometry maximizes the optical interaction in the three-dimensional (3D) structures, while effectively extracting the generated carriers via the conformal PN junction. In this paper, we report characterizations of radial PN junctions that consist of p-type Si micropillars created by deep reactive-ion etching (DRIE) and an n-type layer formed by phosphorus gas diffusion. We use electron-beam induced current (EBIC) microscopy to access the 3D junction profile from the sidewall of the pillars. Our EBIC images reveal uniform PN junctions conformally constructed on the 3D pillar array. Based on Monte-Carlo simulations and EBIC modeling, we estimate local carrier separation/collection efficiency that reflects the quality of the PN junction. We find the EBIC efficiency of the pillar array increases with the incident electron beam energy, consistent with the EBIC behaviors observed in a high-quality planar PN junction. The magnitude of the EBIC efficiency of our pillar array is about 70% at 10?kV, slightly lower than that of the planar device (≈ 81%). We suggest that this reduction could be attributed to the unpassivated pillar surface and the unintended recombination centers in the pillar cores introduced during the DRIE processes. Our results support that the depth-dependent EBIC approach is ideally suitable for evaluating PN junctions formed on micro/nanostructured semiconductors with various geometry.

在制造方面的最新进展使径向结架构具有成本效益和高性能光电器件。与平面PN结不同,径向结的几何形状最大化了三维(3D)结构中的光相互作用,同时有效地通过共形PN结提取生成的载流子。在本文中,我们报告了径向PN结的表征,该结由深反应离子蚀刻(DRIE)产生的p型Si微柱和磷气体扩散形成的n型层组成。我们使用电子束感应电流(EBIC)显微镜从柱的侧壁进入三维结剖面。我们的EBIC图像显示了均匀的PN结在3D柱阵列上共形构造。基于蒙特卡罗模拟和EBIC模型,我们估计了反映PN结质量的局部载流子分离/收集效率。我们发现柱阵的EBIC效率随着入射电子束能量的增加而增加,这与在高质量平面PN结中观察到的EBIC行为一致。柱状阵列的EBIC效率在10?kV,略低于平面器件(≈81%)。我们认为这种减少可能归因于未钝化的矿柱表面和在DRIE过程中引入的矿柱岩心中的意外复合中心。我们的研究结果支持深度依赖EBIC方法非常适合于评估在各种几何形状的微/纳米结构半导体上形成的PN结。
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引用次数: 3
Fine structure of the silk spinning system in the caddisworm, Hydatophylax nigrovittatus (Trichoptera: Limnephilidae) 黑头棘球虫丝系统的精细结构(毛翅目:织丝虫科)
Q3 Immunology and Microbiology Pub Date : 2020-08-06 DOI: 10.1186/s42649-020-00036-5
Hyo-Jeong Kim, Yan Sun, Myung-Jin Moon

Silk is produced by a variety of insects, but only silk made by terrestrial arthropods has been examined in detail. To fill the gap, this study was designed to understand the silk spinning system of aquatic insect. The larvae of caddis flies, Hydatophylax nigrovittatus produce silk through a pair of labial silk glands and use raw silk to protect themselves in the aquatic environment. The result of this study clearly shows that although silk fibers are made under aquatic conditions, the cellular silk production system is quite similar to that of terrestrial arthropods. Typically, silk production in caddisworm has been achieved by two independent processes in the silk glands. This includes the synthesis of silk fibroin in the posterior region, the production of adhesive glycoproteins in the anterior region, which are ultimately accumulated into functional silk dope and converted to a silk ribbon coated with gluey substances. At the cellular level, each substance of fibroin and glycoprotein is specifically synthesized at different locations, and then transported from the rough ER to the Golgi apparatus as transport vesicles, respectively. Thereafter, the secretory vesicles gradually increase in size by vesicular fusion, forming larger secretory granules containing specific proteins. It was found that these granules eventually migrate to the apical membrane and are exocytosed into the lumen by a mechanism of merocrine secretion.

丝是由多种昆虫产生的,但只有陆生节肢动物产生的丝被详细研究过。为了填补这一空白,本研究旨在了解水生昆虫的吐丝系统。海蝇(Hydatophylax nigrovitatus)的幼虫通过一对唇丝腺分泌丝,并利用生丝在水生环境中保护自己。本研究结果清楚地表明,虽然丝纤维是在水生条件下形成的,但细胞产丝系统与陆生节肢动物的产丝系统非常相似。通常,球蚕的产丝是通过两个独立的过程在丝腺中实现的。这包括后区丝素蛋白的合成,前区黏附糖蛋白的产生,最终积聚成功能性丝胶,转化为涂有黏附物质的缎带。在细胞水平上,纤维蛋白和糖蛋白的每一种物质都是在不同的位置特异性合成的,然后分别以运输囊泡的形式从粗内质网转运到高尔基体。此后,分泌囊泡通过囊泡融合逐渐增大,形成含有特定蛋白质的较大分泌颗粒。研究发现,这些颗粒最终迁移到根尖膜,并通过分泌merocrine的机制排出细胞进入管腔。
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引用次数: 0
Replacing critical point drying with a low-cost chemical drying provides comparable surface image quality of glandular trichomes from leaves of Millingtonia hortensis L. f. in scanning electron micrograph 用低成本的化学干燥法代替临界点干燥法,可以获得与之相当的扫描电镜图像质量
Q3 Immunology and Microbiology Pub Date : 2020-07-17 DOI: 10.1186/s42649-020-00035-6
Raktim Bhattacharya, Sulagna Saha, Olga Kostina, Lyudmila Muravnik, Adinpunya Mitra

Sample preparation including dehydration and drying of samples is the most intricate part of scanning electron microscopy. Most current sample preparation protocols use critical-point drying with liquid carbon dioxide. Very few studies have reported samples that were dried using chemical reagents. In this study, we used hexamethyldisilazane, a chemical drying reagent, to prepare plant samples. As glandular trichomes are among the most fragile and sensitive surface structures found on plants, we used Millingtonia hortensis leaf samples as our study materials because they contain abundant glandular trichomes. The results obtained using this new method are identical to those produced via critical-point drying.

样品制备包括样品的脱水和干燥是扫描电子显微镜中最复杂的部分。目前大多数样品制备方案使用液态二氧化碳的临界点干燥。很少有研究报告使用化学试剂干燥样品。在本研究中,我们使用化学干燥试剂六甲基二氮杂烷来制备植物样品。由于腺毛是植物表面结构中最脆弱、最敏感的结构之一,因此我们选择了含有丰富腺毛的密林顿(millington hortensis)叶片样品作为研究材料。用这种新方法得到的结果与临界点干燥得到的结果相同。
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引用次数: 18
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Applied Microscopy
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