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New Advances and Applications in Field-Flow Fractionation. 场流分馏技术的新进展及应用。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091520-052742
Christine L Plavchak, William C Smith, Carmen R M Bria, S Kim Ratanathanawongs Williams

Field-flow fractionation (FFF) is a family of techniques that was created especially for separating and characterizing macromolecules, nanoparticles, and micrometer-sized analytes. It is coming of age as new nanomaterials, polymers, composites, and biohybrids with remarkable properties are introduced and new analytical challenges arise due to synthesis heterogeneities and the motivation to correlate analyte properties with observed performance. Appreciation of the complexity of biological, pharmaceutical, and food systems and the need to monitor multiple components across many size scales have also contributed to FFF's growth. This review highlights recent advances in FFF capabilities, instrumentation, and applications that feature the unique characteristics of different FFF techniques in determining a variety of information, such as averages and distributions in size, composition, shape, architecture, and microstructure and in investigating transformations and function.

场流分馏法(FFF)是一种专门用于分离和表征大分子、纳米颗粒和微米级分析物的技术。随着具有卓越性能的新型纳米材料、聚合物、复合材料和生物杂化材料的引入,由于合成的非均质性以及将分析物性能与观察到的性能相关联的动机,新的分析挑战出现了。对生物、制药和食品系统复杂性的认识,以及对多种规模的多种成分进行监测的需求,也促进了FFF的发展。本文重点介绍了FFF能力、仪器和应用方面的最新进展,这些进展体现了不同FFF技术在确定各种信息(如大小、组成、形状、架构和微观结构的平均值和分布)以及研究转换和功能方面的独特特征。
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引用次数: 8
The Role of Raman Spectroscopy Within Quantitative Metabolomics. 拉曼光谱在定量代谢组学中的作用。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091420-092323
Cassio Lima, Howbeer Muhamadali, Royston Goodacre

Ninety-four years have passed since the discovery of the Raman effect, and there are currently more than 25 different types of Raman-based techniques. The past two decades have witnessed the blossoming of Raman spectroscopy as a powerful physicochemical technique with broad applications within the life sciences. In this review, we critique the use of Raman spectroscopy as a tool for quantitative metabolomics. We overview recent developments of Raman spectroscopy for identification and quantification of disease biomarkers in liquid biopsies, with a focus on the recent advances within surface-enhanced Raman scattering-based methods. Ultimately, we discuss the applications of imaging modalities based on Raman scattering as label-free methods to study the abundance and distribution of biomolecules in cells and tissues, including mammalian, algal, and bacterial cells.

自从拉曼效应被发现以来,已经过去了94年,目前有超过25种不同类型的基于拉曼的技术。在过去的二十年里,拉曼光谱作为一种强大的物理化学技术在生命科学中得到了广泛的应用。在这篇综述中,我们批评使用拉曼光谱作为定量代谢组学的工具。我们概述了拉曼光谱在液体活检中用于疾病生物标志物鉴定和定量的最新进展,重点介绍了基于表面增强拉曼散射的方法的最新进展。最后,我们讨论了基于拉曼散射的成像模式的应用,作为无标记方法来研究细胞和组织中生物分子的丰度和分布,包括哺乳动物、藻类和细菌细胞。
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引用次数: 24
Noncontact Nanoscale Imaging of Cells. 非接触纳米细胞成像。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091420-120101
David Klenerman, Yuri Korchev, Pavel Novak, Andrew Shevchuk

The reduction in ion current as a fine pipette approaches a cell surface allows the cell surface topography to be imaged, with nanoscale resolution, without contact with the delicate cell surface. A variety of different methods have been developed and refined to scan the topography of the dynamic cell surface at high resolution and speed. Measurement of cell topography can be complemented by performing local probing or mapping of the cell surface using the same pipette. This can be done by performing single-channel recording, applying force, delivering agonists, using pipettes fabricated to contain an electrochemical probe, or combining with fluorescence imaging. These methods in combination have great potential to image and map the surface of live cells at the nanoscale.

当细移液管接近细胞表面时,离子电流的减少使细胞表面形貌成像具有纳米级分辨率,而不与微妙的细胞表面接触。各种不同的方法已经被开发和改进,以高分辨率和高速度扫描动态细胞表面的地形。细胞地形的测量可以通过使用相同的移液管进行局部探测或绘制细胞表面来补充。这可以通过执行单通道记录,施加力,输送激动剂,使用制造的含有电化学探针的移液器,或与荧光成像相结合来完成。这些方法的结合在纳米尺度上对活细胞表面进行成像和绘制具有很大的潜力。
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引用次数: 2
In Situ X-Ray Techniques for Electrochemical Interfaces. 电化学界面的原位x射线技术。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091020-100631
Bruna F Baggio, Yvonne Grunder

This article reviews progress in the study of materials using X-ray-based techniques from an electrochemistry perspective. We focus on in situ/in operando surface X-ray scattering, X-ray absorption spectroscopy, and the combination of both methods. The background of these techniques together with key concepts is introduced. Key examples of in situ and in operando investigation of liquid-solid and liquid-liquid interfaces are presented. X-ray scattering and spectroscopy have helped to develop an understanding of the underlying atomic and molecular processes associated with electrocatalysis, electrodeposition, and battery materials. We highlight recent developments, including resonant surface diffraction and time-resolved studies.

本文从电化学的角度综述了基于x射线技术的材料研究进展。我们重点研究了原位/ operando表面x射线散射、x射线吸收光谱以及两种方法的结合。介绍了这些技术的背景和关键概念。介绍了液-固和液-液界面现场和现场研究的关键实例。x射线散射和光谱学有助于理解与电催化、电沉积和电池材料相关的潜在原子和分子过程。我们强调了最近的发展,包括共振表面衍射和时间分辨研究。
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引用次数: 7
Environmental Toxicology Assays Using Organ-on-Chip. 利用器官芯片进行环境毒理学分析。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091620-091335
Patarajarin Akarapipad, Kattika Kaarj, Yan Liang, Jeong-Yeol Yoon

Adverse effects of environmental toxicants to human health have traditionally been assayed using in vitro assays. Organ-on-chip (OOC) is a new platform that can bridge the gaps between in vitro assays (or 3D cell culture) and animal tests. Microenvironments, physical and biochemical stimuli, and adequate sensing and biosensing systems can be integrated into OOC devices to better recapitulate the in vivo tissue and organ behavior and metabolism. While OOCs have extensively been studied for drug toxicity screening, their implementation in environmental toxicology assays is minimal and has limitations. In this review, recent attempts of environmental toxicology assays using OOCs, including multiple-organs-on-chip, are summarized and compared with OOC-based drug toxicity screening. Requirements for further improvements are identified and potential solutions are suggested.

环境毒物对人类健康的不利影响传统上是用体外测定法来测定的。器官芯片(OOC)是一种新的平台,可以弥合体外分析(或3D细胞培养)和动物试验之间的差距。微环境、物理和生化刺激以及足够的传感和生物传感系统可以集成到OOC设备中,以更好地再现体内组织和器官的行为和代谢。虽然OOCs已被广泛用于药物毒性筛选,但它们在环境毒理学分析中的应用很少,而且有局限性。本文综述了近年来使用ooc(包括多器官芯片)进行环境毒理学分析的尝试,并将其与基于ooc的药物毒性筛选进行了比较。确定了进一步改进的需求,并提出了可能的解决方案。
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引用次数: 8
Real-Time Visualization and Monitoring of Physiological Dynamics by Aggregation-Induced Emission Luminogens (AIEgens). 利用聚集诱导发光源(AIEgens)实时可视化和监测生理动力学。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-090420-101149
Xuewen He, Jacky W Y Lam, Ryan T K Kwok, Ben Zhong Tang

Physiological dynamics in living cells and tissues are crucial for maintenance and regulation of their normal activities and functionalities. Tiny fluctuations in physiological microenvironments can leverage significant influences on cell growth, metabolism, differentiation, and apoptosis as well as disease evolution. Fluorescence imaging based on aggregation-induced emission luminogens (AIEgens) exhibits superior advantages in real-time sensing and monitoring of the physiological dynamics in living systems, including its unique properties such as high sensitivity and rapid response, flexible molecular design, and versatile nano- to mesostructural fabrication. The introduction of canonic AIEgens with long-wavelength, near-infrared, or microwave emission, persistent luminescence, and diversified excitation source (e.g., chemo- or bioluminescence) offers researchers a tool to evaluate the resulting molecules with excellent performance in response to subtle fluctuations in bioactivities with broader dimensionalities and deeper hierarchies.

活细胞和组织的生理动力学对于维持和调节其正常活动和功能至关重要。生理微环境的微小波动会对细胞生长、代谢、分化、凋亡以及疾病演变产生重大影响。基于聚集致发射发光源(AIEgens)的荧光成像技术在实时传感和监测生命系统生理动力学方面具有优越的优势,包括高灵敏度和快速响应、灵活的分子设计以及多用途的纳米到介观结构制造。具有长波、近红外或微波发射、持续发光和多样化激发源(如化学或生物发光)的标准AIEgens的引入,为研究人员提供了一种工具,以评估具有优异性能的分子,以响应更广泛维度和更深层次的生物活性的细微波动。
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引用次数: 6
Current Challenges and Recent Developments in Mass Spectrometry-Based Metabolomics. 基于质谱的代谢组学的当前挑战和最新进展。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091620-015205
Stephanie L Collins, Imhoi Koo, Jeffrey M Peters, Philip B Smith, Andrew D Patterson

High-resolution mass spectrometry (MS) has advanced the study of metabolism in living systems by allowing many metabolites to be measured in a single experiment. Although improvements in mass detector sensitivity have facilitated the detection of greater numbers of analytes, compound identification strategies, feature reduction software, and data sharing have not kept up with the influx of MS data. Here, we discuss the ongoing challenges with MS-based metabolomics, including de novo metabolite identification from mass spectra, differentiation of metabolites from environmental contamination, chromatographic separation of isomers, and incomplete MS databases. Because of their popularity and sensitive detection of small molecules, this review focuses on the challenges of liquid chromatography-mass spectrometry-based methods. We then highlight important instrumentational, experimental, and computational tools that have been created to address these challenges and how they have enabled the advancement of metabolomics research.

高分辨率质谱(MS)通过允许在单个实验中测量许多代谢物,促进了对生命系统代谢的研究。虽然质量检测器灵敏度的提高促进了对更多分析物的检测,但化合物鉴定策略、特征约简软件和数据共享并没有跟上质谱数据的涌入。在这里,我们讨论了基于质谱的代谢组学正在面临的挑战,包括从质谱中重新鉴定代谢物,从环境污染中区分代谢物,异构体的色谱分离,以及不完整的质谱数据库。由于液相色谱-质谱技术的普及和对小分子检测的敏感性,本文主要综述了基于液相色谱-质谱技术的方法所面临的挑战。然后,我们重点介绍了为应对这些挑战而创建的重要仪器、实验和计算工具,以及它们如何促进代谢组学研究的进展。
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引用次数: 21
Protein Dynamics by Two-Dimensional Infrared Spectroscopy. 二维红外光谱的蛋白质动力学。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091520-091009
Goran W Tumbic, Md Yeathad Hossan, Megan C Thielges

Proteins function as ensembles of interconverting structures. The motions span from picosecond bond rotations to millisecond and longer subunit displacements. Characterization of functional dynamics on all spatial and temporal scales remains challenging experimentally. Two-dimensional infrared spectroscopy (2D IR) is maturing as a powerful approach for investigating proteins and their dynamics. We outline the advantages of IR spectroscopy, describe 2D IR and the information it provides, and introduce vibrational groups for protein analysis. We highlight example studies that illustrate the power and versatility of 2D IR for characterizing protein dynamics and conclude with a brief discussion of the outlook for biomolecular 2D IR.

蛋白质的功能是相互转换结构的集合体。运动范围从皮秒键旋转到毫秒和更长的亚单元位移。在所有空间和时间尺度上表征功能动力学在实验上仍然具有挑战性。二维红外光谱(2D IR)作为一种研究蛋白质及其动力学的强大方法正在成熟。我们概述了红外光谱的优点,描述了二维红外光谱及其提供的信息,并介绍了用于蛋白质分析的振动基团。我们强调了一些例子研究,说明了二维红外在表征蛋白质动力学方面的能力和多功能性,并简要讨论了生物分子二维红外的前景。
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引用次数: 3
Glycan Labeling and Analysis in Cells and In Vivo. 细胞和体内聚糖的标记和分析。
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091620-091314
Bo Cheng, Qi Tang, Che Zhang, Xing Chen

As one of the major types of biomacromolecules in the cell, glycans play essential functional roles in various biological processes. Compared with proteins and nucleic acids, the analysis of glycans in situ has been more challenging. Herein we review recent advances in the development of methods and strategies for labeling, imaging, and profiling of glycans in cells and in vivo. Cellular glycans can be labeled by affinity-based probes, including lectin and antibody conjugates, direct chemical modification, metabolic glycan labeling, and chemoenzymatic labeling. These methods have been applied to label glycans with fluorophores, which enables the visualization and tracking of glycans in cells, tissues, and living organisms. Alternatively, labeling glycans with affinity tags has enabled the enrichment of glycoproteins for glycoproteomic profiling. Built on the glycan labeling methods, strategies enabling cell-selective and tissue-specific glycan labeling and protein-specific glycan imaging have been developed. With these methods and strategies, researchers are now better poised than ever to dissect the biological function of glycans in physiological or pathological contexts.

聚糖作为细胞内主要的生物大分子之一,在各种生物过程中发挥着重要的功能作用。与蛋白质和核酸相比,聚糖的原位分析更具挑战性。在这里,我们回顾了在细胞和体内聚糖的标记,成像和分析的方法和策略的发展的最新进展。细胞聚糖可以通过基于亲和的探针进行标记,包括凝集素和抗体偶联物、直接化学修饰、代谢聚糖标记和化学酶标记。这些方法已被应用于用荧光团标记聚糖,从而能够可视化和跟踪细胞、组织和生物体中的聚糖。另外,用亲和标签标记聚糖可以使糖蛋白富集用于糖蛋白组学分析。建立在聚糖标记方法的基础上,开发了细胞选择性和组织特异性聚糖标记和蛋白质特异性聚糖成像的策略。有了这些方法和策略,研究人员现在比以往任何时候都更好地剖析聚糖在生理或病理背景下的生物学功能。
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引用次数: 17
Biochemical Sensing with Nanoplasmonic Architectures: We Know How but Do We Know Why? 纳米等离子体结构的生化传感:我们知道如何,但我们知道为什么吗?
Pub Date : 2021-07-27 DOI: 10.1146/annurev-anchem-091420-090751
Andreas Dahlin

Here, the research field of nanoplasmonic sensors is placed under scrutiny, with focus on affinity-based detection using refractive index changes. This review describes how nanostructured plasmonic sensors can deliver unique advantages compared to the established surface plasmon resonance technique, where a planar metal surface is used. At the same time, it shows that these features are actually only useful in quite specific situations. Recent trends in the field are also discussed and some devices that claim extraordinary performance are questioned. It is argued that the most important challenges are related to limited receptor affinity and nonspecific binding rather than instrumental performance. Although some nanoplasmonic sensors may be useful in certain situations, it seems likely that conventional surface plasmon resonance will continue to dominate biomolecular interaction analysis. For detection of analytes in complex samples, plasmonics may be an important tool, but probably not in the form of direct refractometric detection.

本文对纳米等离子体传感器的研究领域进行了深入的探讨,重点是利用折射率变化进行基于亲和力的检测。这篇综述描述了纳米结构等离子体传感器如何与已建立的表面等离子体共振技术相比具有独特的优势,其中使用平面金属表面。同时,它表明这些特性实际上只在非常特定的情况下有用。还讨论了该领域的最新趋势,并对一些声称具有非凡性能的设备提出了质疑。有人认为,最重要的挑战是与有限的受体亲和力和非特异性结合有关,而不是仪器性能。虽然一些纳米等离子体传感器在某些情况下可能是有用的,但似乎传统的表面等离子体共振将继续主导生物分子相互作用的分析。对于复杂样品中的分析物的检测,等离子体可能是一个重要的工具,但可能不是以直接折射检测的形式。
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引用次数: 6
期刊
Annual review of analytical chemistry (Palo Alto, Calif.)
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