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Setting up a light microscopy core facility: Facility design 建立光学显微镜核心设施:设施设计。
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-05-08 DOI: 10.1111/jmi.13301
Timo Zimmermann

The successful operation of a light microscopy core facility depends also on the initial setup of its infrastructure. This article covers the aspects of location selection and room planning and what environmental factors need to be considered. These include light, temperature, vibrations as well as the basic installations needed for microscope operation.

光学显微镜核心设备的成功运行还取决于其基础设施的初始设置。本文将介绍位置选择和房间规划方面的内容,以及需要考虑的环境因素。这些因素包括光线、温度、振动以及显微镜运行所需的基本装置。
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引用次数: 0
High-speed 4-dimensional scanning transmission electron microscopy using compressive sensing techniques 使用压缩传感技术的高速四维扫描透射电子显微镜。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-05-06 DOI: 10.1111/jmi.13315
Alex W. Robinson, Amirafshar Moshtaghpour, Jack Wells, Daniel Nicholls, Miaofang Chi, Ian MacLaren, Angus I. Kirkland, Nigel D. Browning

Here we show that compressive sensing allows 4-dimensional (4-D) STEM data to be obtained and accurately reconstructed with both high-speed and reduced electron fluence. The methodology needed to achieve these results compared to conventional 4-D approaches requires only that a random subset of probe locations is acquired from the typical regular scanning grid, which immediately generates both higher speed and the lower fluence experimentally. We also consider downsampling of the detector, showing that oversampling is inherent within convergent beam electron diffraction (CBED) patterns and that detector downsampling does not reduce precision but allows faster experimental data acquisition. Analysis of an experimental atomic resolution yttrium silicide dataset shows that it is possible to recover over 25 dB peak signal-to-noise ratio in the recovered phase using 0.3% of the total data.

Lay abstract: Four-dimensional scanning transmission electron microscopy (4-D STEM) is a powerful technique for characterizing complex nanoscale structures. In this method, a convergent beam electron diffraction pattern (CBED) is acquired at each probe location during the scan of the sample. This means that a 2-dimensional signal is acquired at each 2-D probe location, equating to a 4-D dataset.

Despite the recent development of fast direct electron detectors, some capable of 100kHz frame rates, the limiting factor for 4-D STEM is acquisition times in the majority of cases, where cameras will typically operate on the order of 2kHz. This means that a raster scan containing 256^2 probe locations can take on the order of 30s, approximately 100-1000 times longer than a conventional STEM imaging technique using monolithic radial detectors. As a result, 4-D STEM acquisitions can be subject to adverse effects such as drift, beam damage, and sample contamination.

Recent advances in computational imaging techniques for STEM have allowed for faster acquisition speeds by way of acquiring only a random subset of probe locations from the field of view. By doing this, the acquisition time is significantly reduced, in some cases by a factor of 10-100 times. The acquired data is then processed to fill-in or inpaint the missing data, taking advantage of the inherently low-complex signals which can be linearly combined to recover the information.

In this work, similar methods are demonstrated for the acquisition of 4-D STEM data, where only a random subset of CBED patterns are acquired over the raster scan. We simulate the compressive sensing acquisition method for 4-D STEM and present our findings for a variety of analysis techniques such as ptychography and differential phase contrast. Our results show that acquisition times can be significantly reduced on the order of 100-300 times, therefore improving existing frame rates, as well as further reducing the electron fluence beyond just using a faster camera.

在这里,我们展示了压缩传感技术可以在高速和低电子通量的条件下获得并精确重建四维(4-D)STEM 数据。与传统的四维方法相比,获得这些结果所需的方法只需要从典型的规则扫描网格中随机获取探针位置子集,这样就能立即在实验中获得更高的速度和更低的电子流。我们还考虑了探测器的下采样,结果表明过采样是会聚束电子衍射(CBED)模式的固有特性,探测器的下采样不会降低精度,反而能加快实验数据的采集。对原子分辨率硅化钇实验数据集的分析表明,只需使用总数据的 0.3%,就可以在恢复相中恢复超过 25 dB 的峰值信噪比。论文摘要:四维扫描透射电子显微镜(4-D STEM)是一种表征复杂纳米级结构的强大技术。在这种方法中,在扫描样品的过程中,每个探针位置都会获得一个会聚束电子衍射图(CBED)。这意味着在每个二维探针位置都能获得一个二维信号,相当于一个四维数据集。尽管最近开发出了快速直接电子探测器,有些探测器的帧频可达 100kHz,但在大多数情况下,4-D STEM 的限制因素是采集时间,摄像机的工作频率通常在 2kHz 左右。这意味着包含 256^2 个探针位置的光栅扫描需要 30 秒左右的时间,比使用单片径向探测器的传统 STEM 成像技术大约长 100-1000 倍。因此,4-D STEM 采集可能会受到漂移、光束损坏和样品污染等不利影响。STEM 计算成像技术的最新进展是,只采集视场中探针位置的随机子集,从而加快了采集速度。通过这种方法,采集时间大大缩短,在某些情况下可缩短 10-100 倍。然后对采集到的数据进行处理,利用固有的低复杂度信号,对缺失的数据进行填充或涂抹,这些信号可以通过线性组合来恢复信息。在这项工作中,我们展示了类似的四维 STEM 数据采集方法,即在光栅扫描中只采集 CBED 图案的随机子集。我们模拟了用于 4-D STEM 的压缩传感采集方法,并介绍了我们对各种分析技术(如层析成像和差分相衬)的研究结果。我们的结果表明,采集时间可以显著缩短 100-300 倍,从而提高现有帧频,并进一步降低电子通量,而不仅仅是使用更快的相机。
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引用次数: 0
Future proofing core facilities with a seven-pillar model 采用七根支柱模式为核心设施的未来发展保驾护航
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-05-03 DOI: 10.1111/jmi.13314
Erin M. Tranfield, Saskia Lippens

Centralised core facilities have evolved into vital components of life science research, transitioning from a primary focus on centralising equipment to ensuring access to technology experts across all facets of an experimental workflow. Herein, we put forward a seven-pillar model to define what a core facility needs to meet its overarching goal of facilitating research. The seven equally weighted pillars are Technology, Core Facility Team, Training, Career Tracks, Technical Support, Community and Transparency. These seven pillars stand on a solid foundation of cultural, operational and framework policies including the elements of transparent and stable funding strategies, modern human resources support, progressive facility leadership and management as well as clear institute strategies and policies. This foundation, among other things, ensures a tight alignment of the core facilities to the vision and mission of the institute. To future-proof core facilities, it is crucial to foster all seven of these pillars, particularly focusing on newly identified pillars such as career tracks, thus enabling core facilities to continue supporting research and catalysing scientific advancement.

Lay abstract: In research, there is a growing trend to bring advanced, high-performance equipment together into a centralised location. This is done to streamline how the equipment purchase is financed, how the equipment is maintained, and to enable an easier approach for research scientists to access these tools in a location that is supported by a team of technology experts who can help scientists use the equipment. These centralised equipment centres are called Core Facilities.

The core facility model is relatively new in science and it requires an adapted approach to how core facilities are built and managed. In this paper, we put forward a seven-pillar model of the important supporting elements of core facilities. These supporting elements are: Technology (the instruments themselves), Core Facility Team (the technology experts who operate the instruments), Training (of the staff and research community), Career Tracks (for the core facility staff), Technical Support (the process of providing help to apply the technology to a scientific question), Community (of research scientist, technology experts and developers) and Transparency (of how the core facility works and the costs associated with using the service). These pillars stand on the bigger foundation of clear policies, guidelines, and leadership approaches at the institutional level. With a focus on these elements, the authors feel core facilities will be well positioned to support scientific discovery in the future.

集中式核心设施已发展成为生命科学研究的重要组成部分,从主要集中设备过渡到确保在实验工作流程的各个方面都能接触到技术专家。在此,我们提出了一个七大支柱模型,以确定核心设施需要哪些条件才能实现其促进研究的总体目标。这七大支柱的权重相同,分别是技术、核心设施团队、培训、职业发展途径、技术支持、社区和透明度。这七大支柱建立在坚实的文化、业务和框架政策基础之上,包括透明和稳定的筹资战略、现代人力资源支持、先进的设施领导和管理以及明确的研究所战略和政策等要素。这一基础,除其他外,确保了核心设施与研究所的愿景和使命紧密结合。要使核心设施面向未来,就必须促进所有这七大支柱的发展,尤其要注重新确定的支柱,如职业发展轨道,从而使核心设施能够继续支持研究工作,推动科学进步。这样做是为了简化设备采购的融资方式、设备的维护方式,并使科研人员能够更容易地在一个由技术专家团队提供支持的地点获得这些工具,从而帮助科学家使用这些设备。这些集中的设备中心被称为核心设施。核心设施模式在科学领域相对较新,需要对核心设施的建设和管理方式进行调整。在本文中,我们提出了核心设施重要支持要素的七根支柱模型。这些支持要素包括技术(仪器本身)、核心设施团队(操作仪器的技术专家)、培训(工作人员和研究团体)、职业发展途径(核心设施工作人员)、技术支持(帮助将技术应用于科学问题的过程)、社区(研究科学家、技术专家和开发人员)和透明度(核心设施如何运作以及使用服务的相关费用)。这些支柱的基础是机构层面的明确政策、指导方针和领导方法。作者认为,有了对这些要素的重视,核心设施就能很好地支持未来的科学发现。
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引用次数: 0
Recognising the importance and impact of Imaging Scientists: Global guidelines for establishing career paths within core facilities 承认成像科学家的重要性和影响力:在核心设施内建立职业发展途径的全球指导方针
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-05-01 DOI: 10.1111/jmi.13307
Graham D. Wright, Kerry A. Thompson, Yara Reis, Johanna Bischof, Philip Edward Hockberger, Michelle S. Itano, Lisa Yen, Stephen Taiye Adelodun, Nikki Bialy, Claire M. Brown, Linda Chaabane, Teng-Leong Chew, Andrew I. Chitty, Fabrice P. Cordelières, Mariana De Niz, Jan Ellenberg, Lize Engelbrecht, Eunice Fabian-Morales, Elnaz Fazeli, Julia Fernandez-Rodriguez, Elisa Ferrando-May, Georgina Fletcher, Graham John Galloway, Adan Guerrero, Jander Matos Guimarães, Caron A. Jacobs, Sachintha Jayasinghe, Eleanor Kable, Gregory T Kitten, Shinya Komoto, Xiaoxiao Ma, Jéssica Araújo Marques, Bryan A. Millis, Kildare Miranda, Peter JohnO'Toole, Sunday Yinka Olatunji, Federica Paina, Cora Noemi Pollak, Clara Prats, Joanna W. Pylvänäinen, Mai Atef Rahmoon, Michael A. Reiche, James Douglas Riches, Andres Hugo Rossi, Jean Salamero, Caroline Thiriet, Stefan Terjung, Aldenora dos Santos Vasconcelos, Antje Keppler

In the dynamic landscape of scientific research, imaging core facilities are vital hubs propelling collaboration and innovation at the technology development and dissemination frontier. Here, we present a collaborative effort led by Global BioImaging (GBI), introducing international recommendations geared towards elevating the careers of Imaging Scientists in core facilities. Despite the critical role of Imaging Scientists in modern research ecosystems, challenges persist in recognising their value, aligning performance metrics and providing avenues for career progression and job security. The challenges encompass a mismatch between classic academic career paths and service-oriented roles, resulting in a lack of understanding regarding the value and impact of Imaging Scientists and core facilities and how to evaluate them properly. They further include challenges around sustainability, dedicated training opportunities and the recruitment and retention of talent. Structured across these interrelated sections, the recommendations within this publication aim to propose globally applicable solutions to navigate these challenges. These recommendations apply equally to colleagues working in other core facilities and research institutions through which access to technologies is facilitated and supported. This publication emphasises the pivotal role of Imaging Scientists in advancing research programs and presents a blueprint for fostering their career progression within institutions all around the world.

在充满活力的科学研究领域,成像核心设施是推动技术开发和传播前沿的合作与创新的重要枢纽。在此,我们介绍由全球生物成像(GBI)领导的一项合作努力,提出了旨在提升核心设施成像科学家职业生涯的国际建议。尽管成像科学家在现代研究生态系统中发挥着至关重要的作用,但在承认他们的价值、调整绩效指标以及提供职业发展和工作保障途径方面,挑战依然存在。这些挑战包括传统的学术职业道路与以服务为导向的角色之间的不匹配,导致人们对成像科学家和核心设施的价值和影响以及如何正确评估他们缺乏了解。这些问题还包括可持续性、专门培训机会以及人才招聘和保留等方面的挑战。在这些相互关联的章节中,本出版物中的建议旨在提出全球适用的解决方案,以应对这些挑战。这些建议同样适用于在其他核心设施和研究机构工作的同事,通过这些设施和机构,可以促进和支持技术的获取。本出版物强调了成像科学家在推进研究计划中的关键作用,并为促进他们在世界各地机构中的职业发展描绘了蓝图。
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引用次数: 0
Suborganellar resolution imaging for the localisation of human glycosylation enzymes in tobacco Golgi bodies 用于烟草高尔基体中人类糖基化酶定位的亚组织细胞分辨率成像技术
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-04-30 DOI: 10.1111/jmi.13311
Alastair J. McGinness, Susan A. Brooks, Richard Strasser, Jennifer Schoberer, Verena Kriechbaumer
Plant cells are a capable system for producing economically and therapeutically important proteins for a variety of applications, and are considered a safer production system than some existing hosts such as bacteria or yeasts. However, plants do not perform protein modifications in the same manner as mammalian cells do. This can impact on protein functionality for plant‐produced human therapeutics. This obstacle can be overcome by creating a plant‐based system capable of ‘humanising’ proteins of interest resulting in a glycosylation profile of synthetic plant‐produced proteins as it would occur in mammalian systems.For this, the human glycosylation enzymes (HuGEs) involved in N‐linked glycosylation N‐acetylglucosaminyltransferase IV and V (GNTIV and GNTV), β‐1,4‐galactosyltransferase (B4GALT1), and α‐2,6‐sialyltransferase (ST6GAL) were expressed in plant cells. For these enzymes to carry out the stepwise glycosylation functions, they need to localise to late Golgi body cisternae. This was achieved by a protein targeting strategy of replacing the mammalian Golgi targeting domains (Cytoplasmic‐Transmembrane‐Stem (CTS) regions) with plant‐specific ones. Using high‐resolution and dynamic confocal microscopy, we show that GNTIV and GNTV were successfully targeted to the medial‐Golgi cisternae while ST6GAL and B4GALT1 were targeted to <jats:italic>trans‐</jats:italic>Golgi cisternae.Plant cells are a promising system to produce human therapeutics for example proteins used in enzyme replacement therapies. Plants can provide safer and cheaper alternatives to existing expression systems such as mammalian cell culture, bacteria or yeast. An important factor for the functionality of therapeutic proteins though are protein modifications specific to human cells. However, plants do not perform protein modifications in the same manner as human cells do. Therefore, plant cells need to be genetically modified to mimic human protein modifications patterns. The modification of importance here, is called N‐linked glycosylation and adds specific sugar molecules onto the proteins.Here we show the expression of four human glycosylation enzymes, which are required for N‐linked glycosylation, in plant cells.In addition, as these protein modifications are carried out in cells resembling a factory production line, it is important that the human glycosylation enzymes be placed in the correct cellular compartments and in the correct order. This is carried out in Golgi bodies. Golgi bodies are composed of several defined stacks termed <jats:italic>cis</jats:italic>‐, medial and <jats:italic>trans</jats:italic>‐Golgi body stacks. For correct protein function, two of these human glycosylation enzymes need to be placed in the medial‐Golgi attacks and the other two in the <jats:italic>trans</jats:italic>‐Golgi stacks. Using high‐resolution laser microscopy in live plant cells, we show here that the human glycosylation enzymes are sent within the cells to the correct Golgi body s
植物细胞是一种能够生产具有经济和治疗意义的蛋白质的系统,可用于多种应用领域,与细菌或酵母等现有宿主相比,植物细胞被认为是一种更安全的生产系统。然而,植物不能像哺乳动物细胞那样进行蛋白质修饰。这可能会影响植物生产的人类治疗药物的蛋白质功能。要克服这一障碍,可以创建一个基于植物的系统,该系统能够对感兴趣的蛋白质进行 "人源化",从而使合成的植物生产的蛋白质具有与哺乳动物系统相同的糖基化特征。为此,在植物细胞中表达了参与N-连接糖基化的人类糖基化酶(HuGEs):N-乙酰葡糖胺基转移酶IV和V(GNTIV和GNTV)、β-1,4-半乳糖基转移酶(B4GALT1)和α-2,6-氨酰基转移酶(ST6GAL)。要使这些酶执行逐步糖基化功能,它们需要定位到高尔基体晚期的小室中。这是通过一种蛋白质靶向策略实现的,即用植物特异性结构域取代哺乳动物高尔基体靶向结构域(细胞质-跨膜-干(CTS)区)。利用高分辨率动态共聚焦显微镜,我们发现 GNTIV 和 GNTV 成功靶向了内侧高尔基细胞,而 ST6GAL 和 B4GALT1 则靶向了反式高尔基细胞。植物细胞是一种很有前途的人类治疗系统,例如酶替代疗法中使用的蛋白质。与哺乳动物细胞培养、细菌或酵母等现有表达系统相比,植物细胞可以提供更安全、更便宜的替代品。不过,影响治疗蛋白质功能的一个重要因素是人体细胞特有的蛋白质修饰。然而,植物进行蛋白质修饰的方式与人体细胞不同。因此,需要对植物细胞进行基因改造,以模仿人类的蛋白质修饰模式。在这里,我们展示了四种人类糖基化酶在植物细胞中的表达情况,这四种酶是进行 N-连接糖基化所必需的。此外,由于这些蛋白质修饰是在类似工厂生产线的细胞中进行的,因此人类糖基化酶必须按照正确的顺序放置在正确的细胞分区中。这项工作在高尔基体中进行。高尔基体由几个确定的堆栈组成,分别称为顺高尔基体堆栈、中高尔基体堆栈和反高尔基体堆栈。为了使蛋白质发挥正确的功能,人类糖基化酶中的两种需要置于内侧-高尔基体堆栈中,另外两种则置于反式-高尔基体堆栈中。我们利用活体植物细胞中的高分辨率激光显微镜,在这里展示了人类糖基化酶在细胞内被送往正确的高尔基体堆栈。这是改造植物细胞以生产人类治疗药物的第一步。
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引用次数: 0
Characterisation and correction of polarisation effects in fluorescently labelled fibres 荧光标记光纤极化效应的表征与校正
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-04-29 DOI: 10.1111/jmi.13308
Nandini Aggarwal, Richard Marsh, Stefania Marcotti, Tanya J Shaw, Brian Stramer, Susan Cox, Siân Culley
SummaryMany biological structures take the form of fibres and filaments, and quantitative analysis of fibre organisation is important for understanding their functions in both normal physiological conditions and disease. In order to visualise these structures, fibres can be fluorescently labelled and imaged, with specialised image analysis methods available for quantifying the degree and strength of fibre alignment. Here we show that fluorescently labelled fibres can display polarised emission, with the strength of this effect varying depending on structure and fluorophore identity. This can bias automated analysis of fibre alignment and mask the true underlying structural organisation. We present a method for quantifying and correcting these polarisation effects without requiring polarisation‐resolved microscopy and demonstrate its efficacy when applied to images of fluorescently labelled collagen gels, allowing for more reliable characterisation of fibre microarchitecture.
摘要许多生物结构都以纤维和细丝的形式存在,对纤维组织的定量分析对于了解它们在正常生理条件和疾病中的功能都非常重要。为了使这些结构可视化,可以对纤维进行荧光标记和成像,并采用专门的图像分析方法来量化纤维排列的程度和强度。在这里,我们展示了荧光标记的纤维可显示极化发射,这种效应的强度因结构和荧光团特性而异。这会使纤维排列的自动分析产生偏差,并掩盖真实的潜在结构组织。我们提出了一种无需偏振分辨显微镜即可量化和校正这些偏振效应的方法,并展示了该方法在应用于荧光标记胶原凝胶图像时的功效,使纤维微观结构的表征更加可靠。
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引用次数: 0
Innovating in a bioimaging core through instrument development 通过仪器开发在生物成像核心进行创新
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-04-29 DOI: 10.1111/jmi.13312
Sebastian Munck, Christof De Bo, Christopher Cawthorne, Julien Colombelli

Developing devices and instrumentation in a bioimaging core facility is an important part of the innovation mandate inherent in the core facility model but is a complex area due to the required skills and investments, and the impossibility of a universally applicable model. Here, we seek to define technological innovation in microscopy and situate it within the wider core facility innovation portfolio, highlighting how strategic development can accelerate access to innovative imaging modalities and increase service range, and thus maintain the cutting edge needed for sustainability. We consider technology development from the perspective of core facility staff and their stakeholders as well as their research environment and aim to present a practical guide to the ‘Why, When, and How’ of developing and integrating innovative technology in the core facility portfolio.

Core facilities need to innovate to stay up to date. However, how to carry out the innovation is not very obvious. One area of innovation in imaging core facilities is the building of optical setups. However, the creation of optical setups requires specific skill sets, time, and investments. Consequently, the topic of whether a core facility should develop optical devices is discussed as controversial. Here, we provide resources that should help get into this topic, and we discuss different options when and how it makes sense to build optical devices in core facilities. We discuss various aspects, including consequences for staff and the relation of the core to the institute, and also broaden the scope toward other areas of innovation.

在生物成像核心设备中开发设备和仪器是核心设备模式固有的创新任务的重要组成部分,但由于所需的技能和投资以及不可能有普遍适用的模式,这是一个复杂的领域。在此,我们试图定义显微镜技术创新,并将其置于更广泛的核心设备创新组合中,强调战略发展如何能够加快创新成像模式的获取,扩大服务范围,从而保持可持续发展所需的尖端优势。我们从核心机构员工及其利益相关者以及研究环境的角度来考虑技术开发问题,旨在为 "为什么、什么时候以及如何 "开发创新技术并将其整合到核心机构组合中提供实用指南。核心设备需要创新,以保持与时俱进。然而,如何进行创新并不十分明显。成像核心设备的创新领域之一是光学装置的建造。然而,创建光学装置需要特定的技能、时间和投资。因此,关于核心设备是否应该开发光学装置的话题一直备受争议。在此,我们提供了有助于了解这一话题的资源,并讨论了在核心设备中制造光学设备的不同选择,以及何时和如何制造光学设备才有意义。我们讨论了各方面的问题,包括对员工的影响以及核心设施与研究所的关系,并将讨论范围扩大到其他创新领域。
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引用次数: 0
Analysis of microscopy techniques to measure segregation in continuous-cast steel slabs 测量连铸钢板偏析的显微镜技术分析
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-04-27 DOI: 10.1111/jmi.13310
Araf Al Rafi, Begoña Santillana, Renfei Feng, Brian G. Thomas, André B. Phillion

The accurate characterisation of centreline segregation requires precise measurements of composition variations over large length scales (101$^{-1}$ m${rm {m}}$) across the centreline of the cast product, while having high resolution, sufficient to quantify the significant composition variations between dendrites due to microsegregation at very small length scales (105m$^{-5}{rm {m}}$). This study investigates the potential of a novel microscopy technique, named Synchrotron Micro X-ray Flurorescence (SMXRF), to generate large-scale high-resolution segregation maps from a steel sample taken from a thin slab caster. Two methods, Point Analysis and Regression Analysis, are proposed for SMXRF data calibration. By comparing with the traditional Laser-Induced Breakdown Spectroscopy (LIBS), and Electron Probe Micro Analyser (EPMA) techniques, we show that SMXRF is successful in quantitative characterisation of centreline segregation. Over large areas (e.g. 12 ×$times$ 16 mm2${rm {mm}}^2$) and at high resolution (10–50 μm$mutext{m}$ pixel size) various techniques yield comparable outcomes in terms of composition maps and solute profiles. The findings also highlight the importance of both high spatial resolution and large field of view to have a quantitative, accurate, and efficient measurement tool to investigate segregation phenomena.

摘要要准确描述中心线偏析的特征,需要精确测量铸件中心线上大长度尺度(10 )的成分变化,同时还要有高分辨率,足以量化由于极小长度尺度的微偏析而导致的树枝状物之间的显著成分变化(10)。本研究调查了一种名为同步辐射微 X 射线荧光(SMXRF)的新型显微镜技术的潜力,该技术可从薄板坯连铸机钢样中生成大尺度高分辨率偏析图。为 SMXRF 数据校准提出了点分析和回归分析两种方法。通过与传统的激光诱导击穿光谱(LIBS)和电子探针显微分析仪(EPMA)技术进行比较,我们发现 SMXRF 成功地对中心线偏析进行了定量表征。在大面积(如 12 16 平方英寸)和高分辨率(10-50 像素大小)条件下,各种技术在成分图和溶质剖面方面产生的结果具有可比性。研究结果还强调了高空间分辨率和大视场对研究偏析现象的定量、准确和高效测量工具的重要性。
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引用次数: 0
Single-shot differential phase contrast microscopy using ring-shaped polarisation multiplexing illumination 使用环形偏振多路复用照明的单次差分相衬显微镜。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-04-25 DOI: 10.1111/jmi.13309
Shengping Wang, Yifu Ma, Mengyuan Xie, Manhong Yao, Zibang Zhang, Jingang Zhong

We propose a differential phase contrast microscopy that enables single-shot phase imaging for unstained biological samples. The proposed approach employs a ring-shaped LED array for polarisation multiplexing illumination and a polarisation camera for image acquisition. As such, multiple images of different polarisation angles can be simultaneously captured with a single shot. Through polarisation demultiplexing, the sample phase can therefore be recovered from the single-shot measurement. Both simulations and experiments demonstrate the effectiveness of the approach. We also demonstrate that ring-shaped illumination enables higher contrast and lower-distortion imaging results than disk-shaped illumination does. The proposed single-shot approach potentially enables phase contrast imaging for live cell samples in vitro.

Lay Description: We propose a microscopy that enables imaging of transparent samples, unstained cells, etc. We demonstrate that the proposed method enables higher contrast and lower-distortion imaging results than conventional methods, and significantly improves imaging efficiency. The proposed method potentially enables dynamic imaging for live cell samples in vitro.

我们提出了一种差分相衬显微镜,可对未染色的生物样本进行单次相位成像。该方法采用环形 LED 阵列进行偏振多路复用照明,并使用偏振相机进行图像采集。因此,一次拍摄就能同时捕捉不同偏振角度的多幅图像。通过偏振解复用,可以从单次测量中恢复样品相位。模拟和实验都证明了这种方法的有效性。我们还证明,与盘状照明相比,环状照明能获得对比度更高、失真度更低的成像结果。所提出的单次方法有可能实现体外活细胞样本的相衬成像。铺层描述:我们提出了一种能对透明样品、未染色细胞等进行成像的显微技术。我们证明,与传统方法相比,所提出的方法能获得对比度更高、失真度更低的成像结果,并能显著提高成像效率。所提出的方法有可能实现体外活细胞样本的动态成像。
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引用次数: 0
Staying on track – Keeping things running in a high-end scientific imaging core facility 保持正常运转 - 确保高端科学成像核心设施的正常运行。
IF 2 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-04-24 DOI: 10.1111/jmi.13304
Oliver Renaud, Nathalie Aulner, Audrey Salles, Nadia Halidi, Maia Brunstein, Adeline Mallet, Karin Aumayr, Stefan Terjung, Daniel Levy, Saskia Lippens, Jean-Marc Verbavatz, Thomas Heuser, Rachel Santarella-Mellwig, Jean-Yves Tinevez, Tatiana Woller, Alexander Botzki, Christopher Cawthorne, The Core4Life Consortium, Sebastian Munck

Modern life science research is a collaborative effort. Few research groups can single-handedly support the necessary equipment, expertise and personnel needed for the ever-expanding portfolio of technologies that are required across multiple disciplines in today's life science endeavours. Thus, research institutes are increasingly setting up scientific core facilities to provide access and specialised support for cutting-edge technologies. Maintaining the momentum needed to carry out leading research while ensuring high-quality daily operations is an ongoing challenge, regardless of the resources allocated to establish such facilities. Here, we outline and discuss the range of activities required to keep things running once a scientific imaging core facility has been established. These include managing a wide range of equipment and users, handling repairs and service contracts, planning for equipment upgrades, renewals, or decommissioning, and continuously upskilling while balancing innovation and consolidation.

现代生命科学研究是一项协作性工作。当今的生命科学研究涉及多个学科,所需的设备、专业知识和人员组合不断扩大,很少有研究小组能够单独提供支持。因此,研究机构正在越来越多地建立科学核心设施,为前沿技术提供获取途径和专业支持。无论为建立此类设施分配了多少资源,在确保高质量日常运作的同时保持开展领先研究的动力都是一项持续的挑战。在此,我们将概述并讨论科学成像核心设施建立后保持正常运行所需的一系列活动。这些活动包括管理各种设备和用户,处理维修和服务合同,规划设备升级、更新或退役,以及在平衡创新和整合的同时不断提高技能。
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引用次数: 0
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Journal of microscopy
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