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Mechanical properties of bone cells studied by atomic force microscopy. 原子力显微镜研究骨细胞的力学特性。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-12-04 DOI: 10.1111/jmi.13373
Xiaoqi Zhang, Zuobin Wang, Haiyue Yu, Zengren Tao, Wei Ji

Osteoblasts are the functional cells capable of bone formation in the bone microenvironment and play an important role in bone growth, development, and the maintenance of bone mass. The cells cultured in vitro are derived from preosteoblasts in tissues and possess the ability to divide and proliferate. Osteoblasts form the bone matrix by secreting collagen and other matrix proteins, which provides a foundation for the deposition of minerals such as calcium and phosphorus, ultimately resulting in the formation of hard bone tissue. Bone diseases affect the quality of life and the aging of the population. Bone diseases such as osteoporosis, fractures, bone tumours, and arthritis have a significant impact on quality of life, especially among the elderly population. These realities remind us that we should pay more attention to bone and joint health. Therefore, it is particularly important to study the imaging and characterisation of mechanical properties of bone cells, which provides a basis for the research of bone diseases in human beings.

成骨细胞是骨微环境中具有成骨功能的细胞,在骨生长发育和骨量维持中起着重要作用。体外培养的细胞来源于组织中的成骨前细胞,具有分裂和增殖的能力。成骨细胞通过分泌胶原蛋白等基质蛋白形成骨基质,为钙、磷等矿物质的沉积提供基础,最终形成坚硬的骨组织。骨病影响生活质量和人口老龄化。骨质疏松症、骨折、骨肿瘤和关节炎等骨病对生活质量有重大影响,特别是在老年人中。这些现实提醒我们,我们应该更加注意骨骼和关节的健康。因此,研究骨细胞力学特性的成像和表征就显得尤为重要,这将为人类骨病的研究提供依据。
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引用次数: 0
Scanning transmission electron tomography to study virus assembly: Review for the retirement of Paul Walther. 用扫描透射电子断层扫描技术研究病毒组装:保罗-瓦尔特退休回顾。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-26 DOI: 10.1111/jmi.13374
Susanne Wieczorek, Jacomina Krijnse Locker

In this short and popular review, we summarise some of our findings analysing the replication cycles of large DNA viruses using scanning transmission electron tomography (STEM tomography) that we applied in the laboratory of Paul Walther. It is also a tribute to a very kind and expert scientist, who recently retired. Transmission electron microscopy (TEM), in particular cryo-EM, has benefited tremendously from recent developments in instrumentation. However, TEM imaging remains limited by the thickness of the specimen and classical thin-section TEM typically generates 2D representations of 3D volumes. Although TEM tomography can partly overcome this limitation, the thickness of the sample, the volume that can be analysed in 3D, remains limiting. STEM tomography can partly overcome this problem, as it allows for the analysis of thicker samples, up to 1 µm in thickness. As such, it is an interesting imaging technique to analyse large DNA viruses, some of which measure 1 µm or more, and which is the focus of our research interest.

在这篇简短而通俗的评论中,我们总结了我们在保罗-瓦尔特(Paul Walther)实验室使用扫描透射电子断层扫描技术(STEM断层扫描)分析大型DNA病毒复制周期的一些研究成果。这也是对最近退休的一位非常和蔼可亲的专家科学家的致敬。透射电子显微镜(TEM),特别是冷冻电子显微镜,从最近的仪器发展中获益匪浅。然而,透射电子显微镜成像仍然受到试样厚度的限制,传统的薄片透射电子显微镜通常生成三维体积的二维图像。虽然 TEM 层析技术可以部分克服这一限制,但样品的厚度,即可进行三维分析的体积,仍然是限制因素。STEM 层析技术可以部分克服这一问题,因为它可以分析厚度达 1 微米的较厚样品。因此,它是一种有趣的成像技术,可用于分析大型 DNA 病毒,其中一些病毒的厚度可达 1 微米或更厚,而这正是我们的研究兴趣所在。
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引用次数: 0
Quantifying whole human hair scalp fibres of varying curl: A micro-computed tomographic study 量化不同卷曲度的整个人类头发头皮纤维:微型计算机断层扫描研究。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-20 DOI: 10.1111/jmi.13365
Claire van den Berg, Nonhlanhla P. Khumalo, Malebogo N. Ngoepe

Scalp hair is a key feature of humans and its variability has been the subject of a broad range of studies. A small subset of these studies has focused on geometric quantification of human scalp hair fibres, however the use of race- and ethnicity-based classification systems makes it challenging to draw objective conclusions about fibre variability. Furthermore, sample preparation techniques for micro-imaging studies often alter the original form of hair fibres. This study sought to determine which of the commonly reported descriptors could be resolved using micro-computed tomography (micro-CT) for fibres of varying curl. Images obtained from micro-CT were used to reconstruct three-dimensional images that were then analysed. The study also explored the capabilities and limitations of micro-CT as an imaging modality by comparing and cross-validating findings with those obtained from scanning electron microscopy (SEM) and laser micrometry. The former deals with surface imaging while the latter deals with cross-sectional measurements. Micro-CT was found to be highly effective at resolving cross-sectional ellipsoidal parameters, but performed more poorly than SEM in reconstructing surface level details at a 2 μm$umutext{m}$ resolution. The technique was, however, able to reveal the presence of the medulla in type VI (high curl) hair fibres. When compared with high curl fibres, greater intra-fibre variability was observed for the low and medium curl fibres, highlighting the importance more objective classification systems.

头皮毛发是人类的一个重要特征,其变异性一直是广泛研究的主题。这些研究中有一小部分侧重于人类头皮毛发纤维的几何量化,但由于使用了基于种族和人种的分类系统,因此很难就纤维的变异性得出客观的结论。此外,用于显微成像研究的样本制备技术往往会改变头发纤维的原始形态。本研究试图确定哪些通常报道的描述符可以通过显微计算机断层扫描(micro-CT)来解析不同卷曲度的纤维。通过微型计算机断层扫描获得的图像被用于重建三维图像,然后对这些图像进行分析。该研究还通过比较和交叉验证扫描电子显微镜(SEM)和激光测微仪获得的结果,探讨了微计算机断层扫描作为一种成像模式的能力和局限性。前者用于表面成像,后者用于横截面测量。研究发现,显微计算机断层扫描在解析横截面椭圆参数方面非常有效,但在以 2 μ m $umutext{m}$ 的分辨率重建表面细节方面的表现比扫描电子显微镜差。不过,该技术能够揭示 VI 型(高卷曲)毛发纤维中髓质的存在。与高卷曲度纤维相比,低卷曲度和中卷曲度纤维的纤维内变异性更大,这凸显了更客观分类系统的重要性。
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引用次数: 0
Diagnostic electron microscopy in human infectious diseases - Methods and applications. 诊断性电子显微镜在人类传染病中的应用--方法与应用。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-19 DOI: 10.1111/jmi.13370
Michael Laue

Diagnostic electron microscopy (EM) is indispensable in all cases of infectious diseases which deserve or profit from the detection of the entire pathogen (i.e. the infectious unit). The focus of its application has shifted during the last decades from routine diagnostics to diagnostics of special cases, emergencies and the investigation of disease pathogenesis. While the focus of application has changed, the methods remain more or less the same. However, since the number of cases for diagnostic EM has declined as the number of laboratories that are able to perform such investigations, the preservation of the present knowledge is important. The aim of this article is to provide a review of the methods and strategies which are useful for diagnostic EM related to infectious diseases in our days. It also addresses weaknesses as well as useful variants or extensions of established methods. The main techniques, negative staining and thin section EM, are described in detail with links to suitable protocols and more recent improvements, such as thin section EM of small volume suspensions. Sample collection, transport and conservation/inactivation are discussed. Strategies of sample examination and requirements for a proper recognition of structures are outlined. Finally, some examples for the actual application of diagnostic EM related to infectious diseases are presented. The outlook section will discuss recent trends in microscopy, such as automated object recognition by machine learning, regarding their potential in supporting diagnostic EM.

诊断性电子显微镜(EM)在所有传染病病例中都是不可或缺的,因为这些病例需要检测整个病原体(即传染单元)或从中获益。在过去的几十年中,电子显微镜的应用重点已从常规诊断转向特殊病例、紧急情况的诊断和疾病发病机制的研究。虽然应用重点发生了变化,但方法却大致相同。不过,由于需要进行电磁诊断的病例数量减少,能够进行此类检查的实验室数量也随之减少,因此保留现有知识非常重要。本文旨在对当今与传染病有关的诊断性电磁学方法和策略进行综述。文章还讨论了现有方法的弱点以及有用的变体或扩展。文章详细介绍了阴性染色和薄片 EM 等主要技术,并链接了合适的方案和最新的改进,如小体积悬浮液的薄片 EM。还讨论了样本的采集、运输和保存/灭活。概述了样品检查策略和正确识别结构的要求。最后,介绍了一些与传染病有关的电磁诊断实际应用实例。展望部分将讨论显微镜的最新趋势,如通过机器学习自动识别物体,以及它们在支持诊断性电磁学方面的潜力。
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引用次数: 0
Brain tissue classification in hyperspectral images using multistage diffusion features and transformer. 利用多级扩散特征和变换器对高光谱图像中的脑组织进行分类
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-19 DOI: 10.1111/jmi.13372
Neetu Sigger, Tuan T Nguyen, Gianluca Tozzi

Brain surgery is a widely practised and effective treatment for brain tumours, but accurately identifying and classifying tumour boundaries is crucial to maximise resection and avoid neurological complications. This precision in classification is essential for guiding surgical decisions and subsequent treatment planning. Hyperspectral (HS) imaging (HSI) is an emerging multidimensional optical imaging method that captures detailed spectral information across multiple wavelengths, allowing for the identification of nuanced differences in tissue composition, with the potential to enhance intraoperative tissue classification. However, current frameworks often require retraining models for each HSI to extract meaningful features, resulting in long processing times and high computational costs. Additionally, most methods utilise the deep semantic features at the end of the network for classification, ignoring the spatial details contained in the shallow features. To overcome these challenges, we propose a novel approach called MedDiffHSI, which combines diffusion and transformer techniques. Our method involves training an unsupervised learning framework based on the diffusion model to extract high-level and low-level spectral-spatial features from HSI. This approach eliminates the need for retraining of spectral-spatial feature learning model, thereby reducing time complexity. We then extract intermediate multistage features from different timestamps for classification using a pretrained denoising U-Net. To fully explore and exploit the rich contextual semantics and textual information hidden in the extracted diffusion feature, we utilise a spectral-spatial attention module. This module not only learns multistage information about features at different depths, but also extracts and enhances effective information from them. Finally, we employ a supervised transformer-based classifier with weighted majority voting (WMV) to perform the HSI classification. To validate our approach, we conduct comprehensive experiments on in vivo brain database data sets and also extend the analysis to include additional HSI data sets for breast cancer to evaluate the framework performance across different types of tissue. The results demonstrate that our framework outperforms existing approaches by using minimal training samples (5%) while achieving state-of-the-art performance.

脑外科手术是治疗脑肿瘤的一种广泛而有效的方法,但要最大限度地切除肿瘤并避免神经系统并发症,准确识别和分类肿瘤边界至关重要。这种精确分类对于指导手术决策和后续治疗计划至关重要。高光谱(HS)成像(HSI)是一种新兴的多维光学成像方法,它能捕捉多个波长的详细光谱信息,从而识别组织成分的细微差别,有望加强术中组织分类。然而,目前的框架通常需要对每个 HSI 重新训练模型,以提取有意义的特征,从而导致处理时间长、计算成本高。此外,大多数方法利用网络末端的深层语义特征进行分类,忽略了浅层特征中包含的空间细节。为了克服这些挑战,我们提出了一种名为 MedDiffHSI 的新方法,它结合了扩散和变换器技术。我们的方法包括训练一个基于扩散模型的无监督学习框架,以从 HSI 中提取高级和低级频谱空间特征。这种方法无需重新训练光谱空间特征学习模型,从而降低了时间复杂性。然后,我们从不同的时间戳中提取中间多级特征,使用预训练的去噪 U-Net 进行分类。为了充分探索和利用所提取的扩散特征中隐藏的丰富的上下文语义和文本信息,我们使用了频谱-空间注意力模块。该模块不仅能学习不同深度特征的多级信息,还能从中提取并增强有效信息。最后,我们采用基于变换器的有监督分类器和加权多数表决(WMV)来进行 HSI 分类。为了验证我们的方法,我们在活体脑部数据库数据集上进行了全面实验,并将分析扩展到乳腺癌的其他 HSI 数据集,以评估框架在不同类型组织中的性能。结果表明,我们的框架只需使用最少的训练样本(5%)就能达到最先进的性能,优于现有的方法。
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引用次数: 0
Novel analysis tool for the distance of gold dimers controlled by the DNA strand length on the DNA origami 通过 DNA 折纸上的 DNA 链长度控制金二聚体距离的新型分析工具。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-15 DOI: 10.1111/jmi.13371
Jannik Guckel, Zhe Liu, Zunhao Wang, Birka Lalkens, Markus Etzkorn, Daesung Park

Metallic nanoparticle dimers have been used to enhance the excitation rate of single-quantum emitters. The interparticle distance (d) of the dimers has a crucial influence on the signal enhancement. Therefore, precise control of d is desired for optimal performance. However, statistical analysis of d has been often restricted to a small number of dimers due to the lack of reliable automatic software tools.

For this reason, we developed a novel analysis tool for automatic dimer analysis. Our approach combines particle detection by circle Hough transformation (CHT) with custom classification routines optimised for distinct types of particles. We applied our tool to scanning electron microscopy (SEM) images and achieved great agreement in dimer detection, reaching an agreement of around 97% between automatic analysis and manual inspection for more than 3000 metallic nanoparticle dimers on DNA origami controlled by a combination of multiple DNA strands.

Our study revealed the effects of the strand length (L) on the distribution of d. Based on geometric consideration, we expected a strong correlation between L and the standard deviation (σ) of d. We could verify this correlation by characterising four dimer designs with different L while analysing more than 1000 dimers per specimen.

金属纳米粒子二聚体已被用于提高单量子发射器的激发率。二聚体的粒子间距(d)对信号增强有至关重要的影响。因此,要想获得最佳性能,就必须精确控制 d。然而,由于缺乏可靠的自动软件工具,对 d 的统计分析往往局限于少数二聚体。为此,我们开发了一种新型分析工具,用于自动二聚体分析。我们的方法将圆圈霍夫变换(CHT)的颗粒检测与针对不同类型颗粒进行优化的定制分类例程相结合。我们将这一工具应用于扫描电子显微镜(SEM)图像,在二聚体检测方面取得了很好的一致性,对于由多条 DNA 链组合控制的 DNA 折纸上的 3000 多个金属纳米粒子二聚体,自动分析与人工检测的一致性达到了 97% 左右。我们的研究揭示了DNA链长度(L)对d分布的影响。基于几何考虑,我们预计L与d的标准偏差(σ)之间存在很强的相关性。
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引用次数: 0
Imaging and observation of microcirculation in bowel mucosa using sidestream dark field imaging 利用侧流暗场成像技术对肠粘膜的微循环进行成像和观察。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-08 DOI: 10.1111/jmi.13367
Keming Jiang, Lihong Chen, Hengyu Zhao, Huanxin Hu, Sicong Lai, Xinzhe Zhao, Hongda Zhang, Jia Ke, Qiongyu Hu

Sidestream dark field (SDF) imaging is a tool for assessing microcirculation, commonly used for early diagnosis and monitoring of sepsis. In this study, we used SDF imaging to observe and assess the microcirculation of the intestinal mucosa during bowel surgery. We also compared different performance between normal mucosa and diseased mucosa using SDF imaging. SDF imaging was conducted in 13 patients to evaluate microcirculation parameters. All patients were assessed at distances of 0, 1, 2, 3 and 4 centimeters (cm) from the edge of the mesentery, respectively. Microcirculatory parameters such as microvascular flow index (MFI), proportion of perfused vessels (PPV), vascular density (VD), total vessel density (TVD), perfused vessel density (PVD) and heterogeneity index (HI) were measured in these patients. Compared to normal intestinal mucosa, the diseased intestinal mucosa exhibited higher values for VD (p = 0.044), TVD (p = 0.006) and PVD (p = 0.007). No significant differences in PPV, MFI and HI were observed between the two groups. The microcirculation parameters (MFI, PPV and PVD) of the intestine at the distal distance of 3 cm were significantly lower than those at a distance of 2 cm (MFI 1.5 (0.75) vs. 3 (0.5), PPV 57.6 (9.1) vs. 97.1 (8.6)% and PVD 11.395 (3.082) vs. 20.726 (4.115) mm/mm2). In conclusion, SDF imaging is an advanced technique that provide real-time visualization of intestinal mucosal microcirculation. It has the potential to assess the blood perfusion of the intestine during surgery.

侧流暗视野(SDF)成像是一种评估微循环的工具,常用于败血症的早期诊断和监测。在这项研究中,我们使用 SDF 成像观察和评估肠道手术中肠粘膜的微循环。我们还利用 SDF 成像比较了正常粘膜和病变粘膜的不同表现。我们对 13 名患者进行了 SDF 成像,以评估微循环参数。所有患者分别在距离肠系膜边缘 0、1、2、3 和 4 厘米(cm)处接受评估。这些患者的微循环参数包括微血管流量指数(MFI)、灌注血管比例(PPV)、血管密度(VD)、总血管密度(TVD)、灌注血管密度(PVD)和异质性指数(HI)。与正常肠粘膜相比,病变肠粘膜的 VD 值(p = 0.044)、TVD 值(p = 0.006)和 PVD 值(p = 0.007)更高。两组间的 PPV、MFI 和 HI 无明显差异。远端距离 3 厘米处的肠道微循环参数(MFI、PPV 和 PVD)明显低于距离 2 厘米处(MFI 1.5 (0.75) vs. 3 (0.5),PPV 57.6 (9.1) vs. 97.1 (8.6)%,PVD 11.395 (3.082) vs. 20.726 (4.115) mm/mm2)。总之,SDF 成像是一种先进的技术,可实时显示肠粘膜微循环。它具有在手术过程中评估肠道血液灌注的潜力。
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引用次数: 0
TOC - Issue Information TOC - 发行信息
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-07 DOI: 10.1111/jmi.13205
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引用次数: 0
Data analysis in imaging (DAIM) – A new RMS science section 成像数据分析(DAIM)--一个新的 RMS 科学部分。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-07 DOI: 10.1111/jmi.13366
Rocco D'Antuono, Laura Murphy, Chas Nelson
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引用次数: 0
In situ isotropic 3D imaging of vasculature perfusion specimens using x-ray microscopic dual-energy CT 利用 X 射线显微双能 CT 对血管灌注标本进行原位各向同性三维成像。
IF 1.5 4区 工程技术 Q3 MICROSCOPY Pub Date : 2024-11-06 DOI: 10.1111/jmi.13369
Stephan Handschuh, Ursula Reichart, Stefan Kummer, Martin Glösmann

Ex vivo x-ray angiography provides high-resolution, three-dimensional information on vascular phenotypes down to the level of capillaries. Sample preparation for ex vivo angiography starts with the removal of blood from the vascular system, followed by perfusion with an x-ray dense contrast agent mixed with a carrier such as gelatine or a polymer. Subsequently, the vascular micro-architecture of harvested organs is imaged in the intact fixed organ. In the present study, we present novel microscopic dual-energy CT (microDECT) imaging protocols that allow to visualise and analyse microvasculature in situ with reference to the morphology of hard and soft tissue. We show that the spectral contrast of µAngiofil and Micropaque barium sulphate in perfused specimens allows for the effective separation of vasculature from mineralised skeletal tissues. Furthermore, we demonstrate the counterstaining of perfused specimens using established x-ray dense contrast agents to depict blood vessels together with the morphology of soft tissue. Phosphotungstic acid (PTA) is used as a counterstain that shows excellent spectral contrast in both µAngiofil and Micropaque barium sulphate–perfused specimens. A novel Sorensen-buffered PTA protocol is introduced as a counterstain for µAngiofil specimens, as the polyurethane polymer is susceptible to artefacts when using conventional staining solutions. Finally, we demonstrate that counterstained samples can be automatically processed into three separate image channels (skeletal tissue, vasculature and stained soft tissue), which offers multiple new options for data analysis. The presented microDECT workflows are suited as tools to screen and quantify microvasculature and can be implemented in various correlative imaging pipelines to target regions of interest for downstream light microscopic investigation.

体外 X 射线血管造影术可提供低至毛细血管水平的高分辨率三维血管表型信息。体外血管造影的样品制备首先要清除血管系统中的血液,然后用一种与明胶或聚合物等载体混合的 X 射线致密造影剂进行灌注。随后,在完整固定的器官中对采集器官的血管微观结构进行成像。在本研究中,我们提出了新颖的显微双能 CT(microDECT)成像方案,可以参照硬组织和软组织的形态,在原位观察和分析微血管。我们的研究表明,µAngiofil 和 Micropaque 硫酸钡在灌注标本中的光谱对比可以有效地将血管从矿化骨骼组织中分离出来。此外,我们还演示了使用成熟的 X 射线致密造影剂对灌注标本进行反染色,以描绘血管和软组织形态。磷钨酸(PTA)作为一种反染色剂,在 µAngiofil 和 Micropaque 硫酸钡灌注标本中都能显示出极佳的光谱对比度。由于聚氨酯聚合物在使用传统染色溶液时容易产生伪影,因此我们引入了一种新颖的索伦森缓冲 PTA 方案作为 µAngiofil 标本的染色剂。最后,我们展示了反染色样本可自动处理成三个独立的图像通道(骨骼组织、血管和染色软组织),这为数据分析提供了多种新选择。所介绍的 microDECT 工作流程适合作为筛选和量化微血管的工具,并可在各种相关成像管道中实施,以瞄准感兴趣的区域进行下游的光学显微镜研究。
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引用次数: 0
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Journal of microscopy
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