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Preliminary study of improved median filter using adaptively mask size in light microscopic image 基于自适应掩模尺寸的改进中值滤波器在微光图像中的初步研究
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfz111
Ji-Youn Kim;Youngjin Lee
This study aimed to develop and evaluate an improved median filter (IMF) with an adaptive mask size for light microscope (LM) images. We acquired images of the mouse first molar using a LM at 100× magnification. The images obtained using our proposed IMF were compared with those from a conventional median filter. Several parameters such as the contrast-to-noise ratio, coefficient of variation, no-reference assessments and peak signal-to-noise ratio were employed to evaluate the image quality quantitatively. The results demonstrated that the proposed IMF could effectively de-noise the LM images and preserve the image details, achieving a better performance than the conventional median filter. This study discusses evaluation of an improved median fi lter with an adaptive mask size for light microscope (LM) images. The results demonstrated that the proposed fi lter could effectively denoise the LM images and preserve the image details, achieving a better performance than the conventional median fi lter.
本研究旨在开发和评估一种用于光学显微镜(LM)图像的具有自适应掩模尺寸的改进中值滤波器(IMF)。我们使用放大100倍的LM获得了小鼠第一磨牙的图像。将使用我们提出的IMF获得的图像与来自传统中值滤波器的图像进行比较。采用对比噪声比、变异系数、无参考评估和峰值信噪比等参数对图像质量进行定量评估。结果表明,所提出的IMF可以有效地对LM图像进行去噪,并保留图像的细节,取得了比传统中值滤波器更好的性能。本研究讨论了一种具有自适应掩模尺寸的改进中值滤波器对光学显微镜(LM)图像的评估。结果表明,该滤波器能够有效地对LM图像进行去噪,保留图像细节,比传统的中值滤波器具有更好的性能。
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引用次数: 5
Visualization of different carrier concentrations in n-type-GaN semiconductors by phase-shifting electron holography with multiple electron biprisms 用多电子双棱镜相移电子全息术实现n型GaN半导体中不同载流子浓度的可视化。
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfz037
Kazuo Yamamoto;Kiyotaka Nakano;Atsushi Tanaka;Yoshio Honda;Yuto Ando;Masaya Ogura;Miko Matsumoto;Satoshi Anada;Yukari Ishikawa;Hiroshi Amano;Tsukasa Hirayama
Phase-shifting electron holography (PS-EH) using a transmission electron microscope (TEM) was applied to visualize layers with different concentrations of carriers activated by Si (at dopant levels of 1019, 1018, 1017 and 1016 atoms cm−3) in n-type GaN semiconductors. To precisely measure the reconstructed phase profiles in the GaN sample, three electron biprisms were used to obtain a series of high-contrast holograms without Fresnel fringes generated by a biprism filament, and a cryo-focused-ion-beam (cryo-FIB) was used to prepare a uniform TEM sample with less distortion in the wide field of view. All layers in a 350-nm-thick TEM sample were distinguished with 1.8-nm spatial resolution and 0.02-rad phase-resolution, and variations of step width in the phase profile (corresponding to depletion width) at the interfaces between the layers were also measured. Thicknesses of the active and inactive layers at each dopant level were estimated from the observed phase profile and the simulation of theoretical band structure. Ratio of active-layer thickness to total thickness of the TEM sample significantly decreased as dopant concentration decreased; thus, a thicker TEM sample is necessary to visualize lower carrier concentrations; for example, to distinguish layers with dopant concentrations of 1016 and 1015 atoms cm−3. It was estimated that sample thickness must be more than 700 nm to make it be possible to detect sub-layers by the combination of PS-EH and cryo-FIB. Phase-shifting electron holography precisely visualized layers with different concentrations of carriers activated by Si (at dopant levels of 1019, 1018, 1017, and 1016 atoms/cm3) in n-GaN semiconductors. A cryo-FIB and triple electron biprisms were used to prepare a uniform TEM sample and to acquire high-contrast holograms without Fresnel fringes.
采用透射电子显微镜(TEM)相移电子全息术(PS-EH)观察了n型GaN半导体中不同载流子浓度的Si激活层(掺杂水平分别为1019、1018、1017和1016原子cm-3)。为了精确测量GaN样品中重构的相位分布,使用三个电子双棱镜获得了一系列由双棱镜灯丝产生的无菲涅耳条纹的高对比度全息图,并使用低温聚焦离子束(cro - fib)制备了宽视场畸变较小的均匀TEM样品。以1.8 nm的空间分辨率和0.02 rad的相位分辨率对350 nm厚TEM样品中的所有层进行了区分,并测量了层间界面处相剖面的阶跃宽度(对应耗尽宽度)的变化。通过观察相分布和理论能带结构的模拟,估计了各掺杂水平上活性层和非活性层的厚度。随着掺杂剂浓度的降低,TEM样品活性层厚度与总厚度之比显著降低;因此,需要较厚的TEM样品来显示较低的载流子浓度;例如,区分掺杂浓度为1016和1015原子cm-3的层。估计样品厚度必须大于700 nm,才能使PS-EH和冷冻fib结合检测子层成为可能。
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引用次数: 2
Theory underpinning multislice simulations with plasmon energy losses 基于等离子体能量损失的多层模拟理论
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfaa003
B G Mendis
The theoretical conditions for small-angle inelastic scattering where the incident electron can effectively be treated as a particle moving in a uniform potential is examined. The motivation for this work is the recent development of a multislice method that combines plasmon energy losses with elastic scattering using Monte Carlo methods. Since plasmon excitation is delocalized, it was assumed that the Bloch wave nature of the incident electron in the crystal does not affect the scattering cross-section. It is shown here that for a delocalized excitation the mixed dynamic form factor term of the scattering cross-section is zero and the scattered intensities follow a Poisson distribution. These features are characteristic of particle-like scattering and validate the use of Monte Carlo methods to model plasmon losses in multislice simulations.
研究了小角度非弹性散射的理论条件,其中入射电子可以有效地被视为在均匀电势中移动的粒子。这项工作的动机是最近开发了一种多层方法,该方法使用蒙特卡罗方法将等离子体能量损失与弹性散射相结合。由于等离子体激元激发是离域的,因此假设晶体中入射电子的布洛赫波性质不会影响散射截面。本文表明,对于离域激发,散射截面的混合动态形状因子项为零,散射强度服从泊松分布。这些特征是类粒子散射的特征,并验证了在多层模拟中使用蒙特卡罗方法对等离子体损失进行建模。
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引用次数: 6
Lensless fourier transform electron holography applied to vortex beam analysis 无透镜傅立叶变换电子全息术在涡旋光束分析中的应用
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfaa008
Ken Harada;Yoshimasa A Ono;Yoshio Takahashi
Lensless Fourier transform holography has been developed. By treating Bragg diffraction waves as object waves and a transmitted spherical wave as a reference wave, these two waves are interfered and recorded as holograms away from the reciprocal plane. In this method, reconstruction of holograms requires only one Fourier transform. Application of this method to analyze vortex beams worked well and their amplitude and phase distributions were obtained on the reciprocal plane. By combining the conventional holography with the developed lensless Fourier transform holography, we can reconstruct and analyze electron waves from the real to reciprocal space continuously.
发展了无透镜傅立叶变换全息术。通过将布拉格衍射波视为物波,将透射球面波视为参考波,这两种波被干涉并记录为远离互易平面的全息图。在这种方法中,全息图的重建只需要一次傅立叶变换。将该方法应用于旋涡光束的分析效果良好,得到了旋涡光束在倒易平面上的振幅和相位分布。将传统的全息术与发展起来的无透镜傅立叶变换全息术相结合,可以连续地重建和分析从实空间到倒易空间的电子波。
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引用次数: 3
Generative and discriminative model-based approaches to microscopic image restoration and segmentation 基于生成和判别模型的显微图像恢复和分割方法
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfaa007
Shin Ishii;Sehyung Lee;Hidetoshi Urakubo;Hideaki Kume;Haruo Kasai
Image processing is one of the most important applications of recent machine learning (ML) technologies. Convolutional neural networks (CNNs), a popular deep learning-based ML architecture, have been developed for image processing applications. However, the application of ML to microscopic images is limited as microscopic images are often 3D/4D, that is, the image sizes can be very large, and the images may suffer from serious noise generated due to optics. In this review, three types of feature reconstruction applications to microscopic images are discussed, which fully utilize the recent advancements in ML technologies. First, multi-frame super-resolution is introduced, based on the formulation of statistical generative model-based techniques such as Bayesian inference. Second, data-driven image restoration is introduced, based on supervised discriminative model-based ML technique. In this application, CNNs are demonstrated to exhibit preferable restoration performance. Third, image segmentation based on data-driven CNNs is introduced. Image segmentation has become immensely popular in object segmentation based on electron microscopy (EM); therefore, we focus on EM image processing.
图像处理是最近机器学习(ML)技术最重要的应用之一。卷积神经网络(CNNs)是一种流行的基于深度学习的ML架构,已被开发用于图像处理应用。然而,ML在微观图像中的应用是有限的,因为微观图像通常是3D/4D,也就是说,图像大小可能非常大,并且图像可能遭受由于光学而产生的严重噪声。在这篇综述中,讨论了三种类型的特征重建在显微图像中的应用,它们充分利用了ML技术的最新进展。首先,在贝叶斯推理等基于统计生成模型的技术的基础上,介绍了多帧超分辨率。其次,介绍了基于监督判别模型的ML技术的数据驱动图像恢复。在该应用中,细胞神经网络被证明具有较好的恢复性能。第三,介绍了基于数据驱动的细胞神经网络的图像分割。图像分割在基于电子显微镜(EM)的对象分割中变得非常流行;因此,我们专注于EM图像处理。
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引用次数: 9
Application of focused ion-beam sampling for sidewall-roughness measurement of free-standing sub-μm objects by atomic force microscopy 聚焦离子束取样在原子力显微镜测量独立亚微米物体侧壁粗糙度中的应用
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfz108
Takaharu Nagatomi;Tatsuya Nakao;Yoko Fujimoto
In the present study, a free-standing object-sampling technique for microelectromechanical systems (MEMS) is developed to measure their sidewall surface roughnesses by atomic force microscopy (AFM). For this purpose, a conventional focused ion beam (FIB) sampling technique widely used for cross-sectional transmission electron microscope specimen preparation was applied. The sub-nm-order roughness parameters were quantitatively measured for sidewalls of Si-bridge test samples. The roughness parameters were compared before and after H2 annealing treatment, which induced smoothing of the surface by migration of the Si atoms. The reduction in the surface roughness by a factor of approximately one-third with 60-s H2 annealing was quantitatively evaluated by AFM. The present study confirms that the developed FIB–AFM technique is one potential approach for quantitatively evaluating the surface-roughness parameters on the oblique faces of free-standing objects in MEMS devices. FIB sampling technique was developed for AFM measurement of side-wall surface roughness of free-standing objects in MEMS devices. We confi rmed that the proposed FIB-AFM technique is one potential and practical approach to quantitatively evaluate surface roughness of oblique faces of free-standing objects in MEMS devices.
在本研究中,开发了一种用于微机电系统(MEMS)的独立物体采样技术,通过原子力显微镜(AFM)测量其侧壁表面粗糙度。为此,应用了广泛用于横截面透射电子显微镜样品制备的传统聚焦离子束(FIB)采样技术。定量测量了硅桥测试样品侧壁的亚纳米级粗糙度参数。比较了H2退火处理前后的粗糙度参数,H2退火处理通过Si原子的迁移使表面光滑。通过AFM定量评估了在60-s H2退火的情况下表面粗糙度降低约三分之一的因子。本研究证实,所开发的FIB–AFM技术是定量评估MEMS器件中独立物体斜面表面粗糙度参数的一种潜在方法。FIB采样技术被开发用于MEMS器件中独立物体侧壁表面粗糙度的AFM测量。我们证实,所提出的FIB-AFM技术是定量评估MEMS器件中独立物体斜面表面粗糙度的一种潜在且实用的方法。
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引用次数: 0
Accuracy improvement of phase estimation in electron holography using noise reduction methods 用降噪方法提高电子全息术相位估计的精度
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfz115
Yoshihiro Midoh;Koji Nakamae
We try to improve the limit of the phase estimation of the interference fringe at low electron dose levels in electron holography by a noise reduction method. In this paper, we focus on unsupervised approaches to apply it to electron beam-sensitive and unknown samples and describe an overview of denoising methods used widely in image processing, such as wiener filter, total variation denoising, nonlocal mean filters and wavelet thresholding. We compare the wavelet hidden Markov model (WHMM) denoising that we have studied so far with the other conventional noise reduction methods. We evaluate the denoise performance of each method using the peak signal-to-noise ratio between noise-free and the target holograms (noisy or denoised holograms) and the root mean-square error (RMSE) between the true phase of the fringe and the measured phase by the discrete Fourier transform phase estimator. We show the denoised holograms for simulation and experimental data by using each noise reduction method and then discuss evaluation indexes obtained from these denoised holograms. From experimental results, it can be seen that the WHMM denoising can reduce the RMSE of fringe phase to about 1/4.5 for noisy simulation holograms and it has stable and good performance for noise reduction of observed holograms with various image qualities.
我们试图通过一种降噪方法来提高电子全息术中低电子剂量水平下干涉条纹相位估计的极限。在本文中,我们重点讨论了将其应用于电子束敏感和未知样本的无监督方法,并概述了在图像处理中广泛使用的去噪方法,如维纳滤波器、全变分去噪、非局部均值滤波器和小波阈值化。我们将迄今为止研究的小波隐马尔可夫模型(WHMM)去噪与其他传统的降噪方法进行了比较。我们使用无噪声和目标全息图(噪声或去噪全息图)之间的峰值信噪比以及条纹的真实相位和离散傅立叶变换相位估计器测量的相位之间的均方根误差(RMSE)来评估每种方法的去噪性能。通过使用各种降噪方法,我们展示了用于模拟和实验数据的降噪全息图,然后讨论了从这些降噪全息图中获得的评估指标。从实验结果可以看出,对于有噪声的模拟全息图,WHMM去噪可以将条纹相位的RMSE降低到约1/4.5,并且对于各种图像质量的观测全息图具有稳定良好的降噪性能。
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引用次数: 8
Automated acquisition of vast numbers of electron holograms with atomic-scale phase information 利用原子尺度相位信息自动获取大量电子全息图
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfaa004
Yoshio Takahashi;Tetsuya Akashi;Atsuko Sato;Toshiaki Tanigaki;Hiroyuki Shinada;Yasukazu Murakami
An automated acquisition system for collecting a large number of electron holograms, to improve the statistical precision of phase analysis, was developed. A technique for shifting the electron beam in combination with stage movement allows data to be acquired over a wide area of a TEM-specimen grid. Undesired drift in the hologram position, which may occur during the hologram acquisition, can be corrected in real time by automated detection of the interference-fringe region in an image. To demonstrate the usefulness of the developed automated hologram acquisition system, gold nanoparticles dispersed on a carbon foil were observed with a 1.2-MV atomic resolution holography electron microscope. The system could obtain 1024 holograms, which provided phase maps for more than 500 nanoparticles with a lateral resolution of 0.14 nm, in just 1 h. The observation results revealed an anomalous increase in mean inner potential for a particle size smaller than 4 nm. The developed automated hologram acquisition system can be applied to improve the precision of phase measurement by averaging many phase images, as demonstrated by single particle analysis for biological entities. Moreover, the system makes it possible to study electrostatic potential of catalysts and other functional nanoparticles at atomic resolution.
为了提高相位分析的统计精度,开发了一种用于收集大量电子全息图的自动采集系统。结合载物台移动来移动电子束的技术允许在TEM样品网格的宽区域上获取数据。可以通过图像中干涉条纹区域的自动检测来实时校正在全息图获取期间可能发生的全息图位置中的不希望的漂移。为了证明所开发的自动全息图采集系统的有用性,用1.2-MV原子分辨率的全息电子显微镜观察了分散在碳箔上的金纳米颗粒。该系统可以在短短1小时内获得1024张全息图,为500多个横向分辨率为0.14 nm的纳米颗粒提供相位图。观察结果显示,对于小于4 nm的颗粒尺寸,平均内电势异常增加。如生物实体的单粒子分析所示,所开发的自动全息图采集系统可以通过对许多相位图像进行平均来提高相位测量的精度。此外,该系统使以原子分辨率研究催化剂和其他功能纳米颗粒的静电势成为可能。
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引用次数: 7
Impaired actin dynamics and suppression of Shank2-mediated spine enlargement in cortactin knockout mice cortactin基因敲除小鼠肌动蛋白动力学受损和Shank2介导的脊柱肥大抑制作用
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfaa001
Shinji Tanaka;Yasutaka Masuda;Akihiro Harada;Shigeo Okabe
Cortactin regulates actin polymerization and stabilizes branched actin network. In neurons, cortactin is enriched in dendritic spines that contain abundant actin polymers. To explore the function of cortactin in dendritic spines, we examined spine morphology and dynamics in cultured neurons taken from cortactin knockout (KO) mice. Histological analysis revealed that the density and morphology of dendritic spines were not significantly different between wild-type (WT) and cortactin KO neurons. Time-lapse imaging of hippocampal slice cultures showed that the extent of spine volume change was similar between WT and cortactin KO neurons. Despite little effect of cortactin deletion on spine morphology and dynamics, actin turnover in dendritic spines was accelerated in cortactin KO neurons. Furthermore, we detected a suppressive effect of cortactin KO on spine head size under the condition of excessive spine enlargement induced by overexpression of a prominent postsynaptic density protein Shank2. These results suggest that cortactin may have a role in maintaining actin organization by stabilizing actin filaments near the postsynaptic density. Cortactin is an actin-binding protein enriched in the synapse. Neurons without cortactin show accelerated spine actin turnover and reduced ability to increase spine size triggered by overexpression of Shank2, a prominent postsynaptic protein. Cortactin may play a role in actin fi lament maintenance and spine shape regulation in mammalian neurons.
Cortactin调节肌动蛋白聚合并稳定支链肌动蛋白网络。在神经元中,cortactin在树突棘中富集,树突棘中含有丰富的肌动蛋白聚合物。为了探索cortactin在树突棘中的功能,我们检测了cortactin敲除(KO)小鼠培养神经元的棘形态和动力学。组织学分析显示,野生型(WT)和cortactin KO神经元的树突棘密度和形态没有显著差异。海马切片培养的延时成像显示,WT和cortactin KO神经元的脊柱体积变化程度相似。尽管cortactin缺失对棘的形态和动力学影响很小,但在cortactin KO神经元中,树突棘的肌动蛋白周转加速。此外,我们检测到在突触后密度蛋白Shank2过表达诱导的过度脊柱增大的情况下,cortactin KO对脊柱头部大小的抑制作用。这些结果表明,cortactin可能通过稳定突触后密度附近的肌动蛋白丝来维持肌动蛋白组织。Cortactin是一种在突触中富集的肌动蛋白结合蛋白。不含cortactin的神经元表现出由Shank2(一种重要的突触后蛋白)过表达引发的脊椎肌动蛋白周转加速和增加脊椎大小的能力降低。Cortactin可能在哺乳动物神经元的肌动蛋白丝维持和脊柱形状调节中发挥作用。
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引用次数: 5
Energy-dispersive X-ray spectroscopy for an atomic-scale quantitative analysis of Pd–Pt core-shell nanoparticles 用于原子尺度定量分析Pd–Pt核壳纳米颗粒的能量色散X射线光谱
IF 1.8 4区 工程技术 Pub Date : 2019-11-01 DOI: 10.1093/jmicro/dfz113
Shin Inamoto;Yuji Otsuka
The properties of core-shell nanoparticles, which are used for many catalytic processes as an alternative to platinum, depend on the size of both the particle and the shell. It is thus necessary to develop a quantitative method to determine the shell thickness. Pd–Pt core-shell particles were analyzed using scanning transmission electron microscopy (STEM) and energy-dispersive X-ray spectroscopy (EDX). Quantitative EDX line profiles acquired from the core-shell particle were compared to four core-shell models. The results indicate that the thickness of the Pt shell corresponds to two atomic layers. Meanwhile, high-angle annular dark-field STEM images from the same particle were analyzed and compared to simulated images. Again, this experiment demonstrates that the shell thickness was of two atomic layers. Our results indicate that, in small particles, it is possible to use EDX for a precise atomic-scale quantitative analysis. This article discusses the quantifi cation of EDX map acquired from Pd-Pt core-shell nanoparticles at the atomic scale. The EDX analysis provides that the thickness of the Pt shell corresponds to two atomic layers. The result indicates EDX is a great tool when studying core-shell nanoparticles.
核壳纳米颗粒作为铂的替代品用于许多催化过程,其性质取决于颗粒和外壳的大小。因此,有必要开发一种定量方法来确定壳体厚度。使用扫描透射电子显微镜(STEM)和能量色散X射线光谱(EDX)分析了Pd–Pt核壳颗粒。将从核壳颗粒获得的定量EDX谱线与四个核壳模型进行比较。结果表明,Pt壳层的厚度对应于两个原子层。同时,分析了来自同一粒子的高角度环形暗场STEM图像,并将其与模拟图像进行了比较。这个实验再次证明了壳层的厚度是两个原子层。我们的结果表明,在小颗粒中,可以使用EDX进行精确的原子级定量分析。本文讨论了从Pd-Pt核壳纳米粒子中获得的EDX图谱在原子尺度上的量子化。EDX分析提供了Pt壳的厚度对应于两个原子层。结果表明EDX是研究核壳纳米粒子的一个很好的工具。
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引用次数: 3
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