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Development of Simple and Rapid Bead-Based Cytometric Immunoassays Using Superparamagnetic Hybrid Core–Shell Microparticles 利用超顺磁性混合核壳微粒开发简单快速的珠基细胞免疫测定方法
Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-17 DOI: 10.1021/acsmeasuresciau.4c00038
Charlie Tobias, Daniel López-Puertollano, Antonio Abad-Somovilla, Josep V. Mercader, Antonio Abad-Fuentes, Knut Rurack
Flow cytometry-based immunoassays are valuable in biomedical research and clinical applications due to their high throughput and multianalyte capability, but their adoption in areas such as food safety and environmental monitoring is limited by long assay times and complex workflows. Rapid, simplified bead-based cytometric immunoassays are needed to make these methods viable for point-of-need applications, especially with the increasing accessibility of miniaturized cytometers. This work introduces superparamagnetic hybrid polystyrene-silica core–shell microparticles as promising alternatives to conventional polymer beads in competitive cytometric immunoassays. These beads, featuring high specificity, sensitivity, and excellent handling capabilities via magnetic separation, were evaluated with three different antibodies and binding methods, showing variations in signal intensity based on the antibody and its attachment method. The optimal performance was achieved through a secondary antibody binding approach, providing strong and consistent signals with minimal uncertainty. The optimized protocol made it possible to achieve a detection limit of 0.025 nM in a total assay time of only 15 min and was successfully used to detect ochratoxin A (OTA) in raw flour samples. This work highlights the potential of these beads as versatile tools for flow cytometry-based immunoassays, with significant implications for food safety, animal health, environmental monitoring, and clinical diagnostics.
基于流式细胞仪的免疫测定因其高通量和多分析能力而在生物医学研究和临床应用中具有重要价值,但其在食品安全和环境监测等领域的应用却受到化验时间长和工作流程复杂的限制。需要快速、简化的基于微珠的细胞计量免疫测定,使这些方法在需求点应用中切实可行,特别是随着微型细胞仪的日益普及。这项工作介绍了超顺磁性混合聚苯乙烯-二氧化硅核壳微粒,作为竞争性细胞免疫测定中传统聚合物微珠的有前途的替代品。这些珠子具有高特异性、高灵敏度和通过磁性分离的出色处理能力,我们用三种不同的抗体和结合方法对其进行了评估,结果表明,根据抗体及其附着方法的不同,信号强度也有差异。通过二抗结合的方法实现了最佳性能,可提供强而稳定的信号,且不确定性最小。优化后的方案使检测限达到了 0.025 nM,总检测时间仅为 15 分钟,并成功用于检测生面粉样品中的赭曲霉毒素 A(OTA)。这项工作凸显了这些微珠作为流式细胞仪免疫测定多功能工具的潜力,对食品安全、动物健康、环境监测和临床诊断具有重要意义。
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引用次数: 0
Colorimetric Hybridization Sensor for DNA Mimic of a SARS-CoV-2 RNA Marker: Direct and Inverse Bioanalysis 用于 SARS-CoV-2 RNA 标记 DNA 模拟的比色杂交传感器:直接和逆向生物分析
Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-09 DOI: 10.1021/acsmeasuresciau.4c00043
Zia ul Quasim Syed, Sathya Samaraweera, Zhuo Wang, James Kelby Schrader, Colton Scott, Joshua Schut, Dozier Johnson Smith, Joshua D. Ramsey, Sadagopan Krishnan
This article presents a colorimetric visual biosensor designed for direct application in undiluted biofluids, which holds significant promise for point-of-need applications. Unlike traditional biosensors that struggle with heavily diluted sample matrices, the presented biosensor does not require any instrumentation or trained personnel, making it highly practical. The sensor features an oligonucleotide probe covalently attached to magnetically separable magnetite (Fe3O4) particles. This probe selectively captures a DNA mimic of the SARS-CoV-2 RNA sequence via a base-pair hybridization. The DNA mimic oligomer sequence was tested in a buffer solution, undiluted serum, and undiluted salivary biofluids. A second complementary hybridization sequence with a biotin tag was used to bind the target oligomer already hybridized to the magnetic particle-conjugated capture probe. Subsequent detection of the target oligomer was accomplished through high-affinity selective binding of streptavidin-peroxidase labels with the detection probe biotin units for visual colorimetric detection in the presence of 3,3′,5,5′-tetramethylbenzidine and hydrogen peroxide. Inverse assaying of the unbound-free streptavidin-peroxidase labels left in the detection reagent solution offered a reverse trend to the target oligomer concentration, as anticipated. We obtained detection limits of 1 fM (buffer assay), 1 pM (undiluted serum assay), and 1 pM (undiluted saliva assay) and with the linear ranges of 1 fM–10 nM (buffer assay), 1 pM–1 nM (undiluted serum assay), and 1 pM–1 nM (undiluted saliva assay), respectively. The assays in different biofluids allowed for the estimation of the analytical performance and the effect of sample matrices on the detection limits and calibration sensitivity.
本文介绍了一种可直接应用于未稀释生物流体的比色视觉生物传感器,它在需求点应用方面前景广阔。传统的生物传感器在严重稀释的样品基质中难以发挥作用,而本文介绍的生物传感器则不同,它不需要任何仪器或训练有素的人员,因此非常实用。该传感器将寡核苷酸探针共价连接到磁性可分离的磁铁矿(Fe3O4)颗粒上。该探针通过碱基对杂交选择性地捕获 SARS-CoV-2 RNA 序列的 DNA 模拟物。在缓冲溶液、未稀释的血清和未稀释的唾液生物流体中测试了 DNA 模拟寡聚体序列。第二个带有生物素标记的互补杂交序列用于结合已经与磁性颗粒结合的捕获探针杂交的目标寡聚体。在 3,3′,5,5′-四甲基联苯胺和过氧化氢存在的情况下,通过链霉亲和素-过氧化物酶标签与检测探针生物素单位的高亲和性选择性结合进行可视比色检测,从而实现对目标寡聚体的后续检测。对检测试剂溶液中残留的未结合的链霉亲和素-过氧化物酶标签进行反向测定,结果与预期的目标低聚物浓度呈反向趋势。我们得到的检测限分别为 1 fM(缓冲液检测)、1 pM(未稀释血清检测)和 1 pM(未稀释唾液检测),线性范围分别为 1 fM-10 nM(缓冲液检测)、1 pM-1 nM(未稀释血清检测)和 1 pM-1 nM(未稀释唾液检测)。在不同的生物流体中进行测定,可以评估分析性能以及样品基质对检测限和校准灵敏度的影响。
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引用次数: 0
Computer-Assisted Processing of Current Step Signals in Single Blocking Impact Electrochemistry 计算机辅助处理单阻断影响电化学中的电流阶跃信号
Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-06 DOI: 10.1021/acsmeasuresciau.4c00046
Arthur Langlard, Hassiba Smida, Romain Chevalet, Christine Thobie-Gautier, Mohammed Boujtita, Estelle Lebègue
Current step signals related to single-entity collisions in blocking impact electrochemistry were analyzed by computer-assisted processing for estimating the size distributions of various particles. In this work, three different types of entities were studied by single blocking impact electrochemistry: polystyrene nanospheres (350 nm diameter) and microspheres (1 μm diameter), phospholipid liposomes (300 nm diameter) and two different strains of Gram-negative bacillus bacteria (Escherichia coli and Shewanella oneidensis). The size estimations of these different entities from the current step signal analysis were compared and discussed according to the shape and size of each entity. From the magnitude of the current step transient, the size distribution of each entity was calculated by a new computer program assisting in the detection and analysis of single impact events in chronoamperometry measurements. The data processing showed that the size distributions obtained from the electrochemical data agreed with the dynamic light scattering and atomic force microscopy data for nanospheres and liposomes. In contrast, the size estimation calculated from the electrochemical data was underestimated for microspheres and bacteria. We demonstrated that our computer program was efficient for detecting and analyzing the collision events in single blocking impact electrochemistry for various entities from spherical hard nanoparticles to micrometer-sized rod-shaped living bacteria.
通过计算机辅助处理,分析了阻断冲击电化学中与单实体碰撞有关的电流阶跃信号,以估计各种粒子的尺寸分布。在这项工作中,通过单次阻塞撞击电化学研究了三种不同类型的实体:聚苯乙烯纳米球(直径为 350 nm)和微球(直径为 1 μm)、磷脂脂质体(直径为 300 nm)以及两种不同的革兰氏阴性杆菌菌株(大肠杆菌和 Shewanella oneidensis)。根据每个实体的形状和大小,对电流阶跃信号分析得出的这些不同实体的大小估计进行了比较和讨论。根据电流阶跃瞬态的大小,用一个新的计算机程序计算出了每个实体的大小分布,该程序可协助检测和分析计时器振荡测量中的单次冲击事件。数据处理结果表明,从电化学数据中获得的尺寸分布与纳米球和脂质体的动态光散射和原子力显微镜数据一致。相反,对于微球和细菌,根据电化学数据计算出的粒度估计值被低估了。我们证明了我们的计算机程序可以有效地检测和分析从球形硬质纳米粒子到微米大小的杆状活细菌等各种实体在单阻挡撞击电化学中的碰撞事件。
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引用次数: 0
Super-Resolution Microscopic Imaging of Lipid Droplets in Living Cells via Carbonized Polymer Dot-Based Polarity-Responsive Nanoprobe 通过基于碳化聚合物点的极性响应纳米探针对活细胞中的脂滴进行超分辨率显微成像
Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-06 DOI: 10.1021/acsmeasuresciau.4c00049
Zepeng Huo, Zitong Yu, Weiqing Xu, Shuping Xu
Lipid droplets (LDs) are dynamic subcellular organelles that participate in various physiological processes, and their abnormality can also lead to various diseases. Tracing the dynamics of LDs in living cells will be valuable for understanding cell physiological states. Here, we employed a structured light illumination super-resolution imaging assisted with a carbonized polymer dot (CPD)-based fluorescence nanoprobe to track the travel paths of LDs and other organelles. The CPDs we developed are highly biocompatible with living cells and exhibit a highly sensitive response to solvent polarity, allowing for high specificity in staining LDs in living cells. Aided by these nanoprobes, we successfully observed many real-time LD-involved dynamics in living cells, such as intracellular LD interactions, communications with other organelles, and dynamic behaviors under external stimuli (oxidative stress inducer). These studies deepen our comprehension of the physiological role of LDs and drive the advancement of super-resolution fluorescent probes.
脂滴(LDs)是参与各种生理过程的动态亚细胞器,其异常也会导致各种疾病。追踪活细胞中脂滴的动态变化对了解细胞生理状态很有价值。在这里,我们采用了一种结构光照明超分辨率成像技术,辅助以碳化聚合物点(CPD)为基础的荧光纳米探针来跟踪 LDs 和其他细胞器的移动路径。我们开发的碳化聚合物点与活细胞具有高度的生物相容性,并对溶剂极性表现出高度敏感的反应,因此可对活细胞中的低密度脂蛋白进行高特异性染色。在这些纳米探针的帮助下,我们成功地观察到了活细胞中许多与 LD 有关的实时动态,如细胞内 LD 的相互作用、与其他细胞器的通讯以及在外部刺激(氧化应激诱导剂)下的动态行为。这些研究加深了我们对 LD 生理作用的理解,推动了超分辨率荧光探针的发展。
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引用次数: 0
Super-Resolution Microscopic Imaging of Lipid Droplets in Living Cells via Carbonized Polymer Dot-Based Polarity-Responsive Nanoprobe 通过基于碳化聚合物点的极性响应纳米探针对活细胞中的脂滴进行超分辨率显微成像
IF 4.6 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-06 DOI: 10.1021/acsmeasuresciau.4c0004910.1021/acsmeasuresciau.4c00049
Zepeng Huo, Zitong Yu, Weiqing Xu and Shuping Xu*, 

Lipid droplets (LDs) are dynamic subcellular organelles that participate in various physiological processes, and their abnormality can also lead to various diseases. Tracing the dynamics of LDs in living cells will be valuable for understanding cell physiological states. Here, we employed a structured light illumination super-resolution imaging assisted with a carbonized polymer dot (CPD)-based fluorescence nanoprobe to track the travel paths of LDs and other organelles. The CPDs we developed are highly biocompatible with living cells and exhibit a highly sensitive response to solvent polarity, allowing for high specificity in staining LDs in living cells. Aided by these nanoprobes, we successfully observed many real-time LD-involved dynamics in living cells, such as intracellular LD interactions, communications with other organelles, and dynamic behaviors under external stimuli (oxidative stress inducer). These studies deepen our comprehension of the physiological role of LDs and drive the advancement of super-resolution fluorescent probes.

脂滴(LDs)是参与各种生理过程的动态亚细胞器,其异常也会导致各种疾病。追踪活细胞中脂滴的动态变化对了解细胞生理状态很有价值。在这里,我们采用了一种结构光照明超分辨率成像技术,辅助以碳化聚合物点(CPD)为基础的荧光纳米探针来跟踪 LDs 和其他细胞器的移动路径。我们开发的碳化聚合物点与活细胞具有高度的生物相容性,对溶剂极性的反应非常灵敏,可对活细胞中的LDs进行高特异性染色。在这些纳米探针的帮助下,我们成功地观察到了活细胞中许多与 LD 有关的实时动态,如细胞内 LD 的相互作用、与其他细胞器的通讯以及在外部刺激(氧化应激诱导剂)下的动态行为。这些研究加深了我们对 LD 生理作用的理解,推动了超分辨率荧光探针的发展。
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引用次数: 0
Computer-Assisted Processing of Current Step Signals in Single Blocking Impact Electrochemistry 计算机辅助处理单阻断影响电化学中的电流阶跃信号
IF 4.6 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-06 DOI: 10.1021/acsmeasuresciau.4c0004610.1021/acsmeasuresciau.4c00046
Arthur Langlard, Hassiba Smida, Romain Chevalet, Christine Thobie-Gautier, Mohammed Boujtita and Estelle Lebègue*, 

Current step signals related to single-entity collisions in blocking impact electrochemistry were analyzed by computer-assisted processing for estimating the size distributions of various particles. In this work, three different types of entities were studied by single blocking impact electrochemistry: polystyrene nanospheres (350 nm diameter) and microspheres (1 μm diameter), phospholipid liposomes (300 nm diameter) and two different strains of Gram-negative bacillus bacteria (Escherichia coli and Shewanella oneidensis). The size estimations of these different entities from the current step signal analysis were compared and discussed according to the shape and size of each entity. From the magnitude of the current step transient, the size distribution of each entity was calculated by a new computer program assisting in the detection and analysis of single impact events in chronoamperometry measurements. The data processing showed that the size distributions obtained from the electrochemical data agreed with the dynamic light scattering and atomic force microscopy data for nanospheres and liposomes. In contrast, the size estimation calculated from the electrochemical data was underestimated for microspheres and bacteria. We demonstrated that our computer program was efficient for detecting and analyzing the collision events in single blocking impact electrochemistry for various entities from spherical hard nanoparticles to micrometer-sized rod-shaped living bacteria.

通过计算机辅助处理,分析了阻断冲击电化学中与单实体碰撞有关的电流阶跃信号,以估计各种粒子的尺寸分布。在这项工作中,通过单次阻塞撞击电化学研究了三种不同类型的实体:聚苯乙烯纳米球(直径为 350 nm)和微球(直径为 1 μm)、磷脂脂质体(直径为 300 nm)以及两种不同的革兰氏阴性杆菌菌株(大肠杆菌和 Shewanella oneidensis)。根据每个实体的形状和大小,对电流阶跃信号分析得出的这些不同实体的大小估计进行了比较和讨论。根据电流阶跃瞬态的大小,用一个新的计算机程序计算出了每个实体的大小分布,该程序可协助检测和分析计时器振荡测量中的单次冲击事件。数据处理结果表明,从电化学数据中获得的尺寸分布与纳米球和脂质体的动态光散射和原子力显微镜数据一致。相反,对于微球和细菌,根据电化学数据计算出的粒度估计值被低估了。我们证明了我们的计算机程序可以有效地检测和分析从球形硬质纳米粒子到微米大小的杆状活细菌等各种实体在单阻挡撞击电化学中的碰撞事件。
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引用次数: 0
Simultaneous Imaging of Temperature and Oxygen by Utilizing Thermally Activated Delayed Fluorescence and Phosphorescence of a Single Indicator 利用单一指示剂的热激活延迟荧光和磷光同时成像温度和氧气
IF 4.6 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-05 DOI: 10.1021/acsmeasuresciau.4c0004010.1021/acsmeasuresciau.4c00040
Georg Schwendt, Andrey V. Kalinichev, Sergey M. Borisov* and Klaus Koren*, 

Chemical gradients are essential in biological systems, affecting processes like microbial activity in soils and nutrient cycling. Traditional tools, such as microsensors, offer high-resolution data but are limited to one-dimensional measurements. Planar optodes allow for two-dimensional (2D) and three-dimensional (3D) chemical imaging but are often sensitive to temperature changes. This study presents an advanced dual-emission optical sensor that simultaneously measures temperature and oxygen using a modified platinum(II) meso-tetrakis(3,5-ditert-butylphenyl)-tetra(2-tert-butyl-1,4-naphthoquinono)porphyrin. The ratio between thermally activated delayed fluorescence and phosphorescence was optimized by modifying platinum(II) naphthoquinonoporphyrin with tert-butyl groups which simultaneously improved solubility in apolar solvents and polymer matrix (polystyrene). This dual-function sensor enables two-parameter chemical imaging with a consumer-grade RGB camera or a hyperspectral camera. We demonstrated 2D visualization of temperature and oxygen distribution in a model soil system. The RGB camera provided rapid and cost-effective imaging, while the hyperspectral camera offered more detailed spectral information despite some limitations. Our findings revealed the formation of a stable temperature gradient and oxygen depletion, driven by water content and temperature-sensitive microbial activity. This dual O2/T sensor, with further potential improvements, shows considerable promise for advanced multiparameter sensing in complex biological and environmental studies, providing deeper insights into dynamic microenvironments.

化学梯度在生物系统中至关重要,影响着土壤中微生物活动和养分循环等过程。微传感器等传统工具可提供高分辨率数据,但仅限于一维测量。平面光电二极管可进行二维(2D)和三维(3D)化学成像,但通常对温度变化很敏感。本研究提出了一种先进的双发射光学传感器,利用改性铂(II)介-四(3,5-二叔丁基苯基)-四(2-叔丁基-1,4-萘醌)卟啉同时测量温度和氧气。通过对铂(II)萘醌卟啉进行叔丁基修饰,优化了热激活延迟荧光和磷光之间的比例,同时提高了在无极性溶剂和聚合物基质(聚苯乙烯)中的溶解度。这种双功能传感器可通过消费级 RGB 相机或高光谱相机实现双参数化学成像。我们演示了模型土壤系统中温度和氧气分布的二维可视化。RGB 相机能快速、经济地进行成像,而高光谱相机尽管有一些局限性,但能提供更详细的光谱信息。我们的研究结果表明,在含水量和对温度敏感的微生物活动的驱动下,形成了稳定的温度梯度和氧气耗竭。这种 O2/T 双传感器具有进一步改进的潜力,在复杂的生物和环境研究中显示出先进的多参数传感技术的巨大前景,为动态微环境提供了更深入的洞察力。
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引用次数: 0
Simultaneous Imaging of Temperature and Oxygen by Utilizing Thermally Activated Delayed Fluorescence and Phosphorescence of a Single Indicator 利用单一指示剂的热激活延迟荧光和磷光同时成像温度和氧气
Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-09-05 DOI: 10.1021/acsmeasuresciau.4c00040
Georg Schwendt, Andrey V. Kalinichev, Sergey M. Borisov, Klaus Koren
Chemical gradients are essential in biological systems, affecting processes like microbial activity in soils and nutrient cycling. Traditional tools, such as microsensors, offer high-resolution data but are limited to one-dimensional measurements. Planar optodes allow for two-dimensional (2D) and three-dimensional (3D) chemical imaging but are often sensitive to temperature changes. This study presents an advanced dual-emission optical sensor that simultaneously measures temperature and oxygen using a modified platinum(II) meso-tetrakis(3,5-ditert-butylphenyl)-tetra(2-tert-butyl-1,4-naphthoquinono)porphyrin. The ratio between thermally activated delayed fluorescence and phosphorescence was optimized by modifying platinum(II) naphthoquinonoporphyrin with tert-butyl groups which simultaneously improved solubility in apolar solvents and polymer matrix (polystyrene). This dual-function sensor enables two-parameter chemical imaging with a consumer-grade RGB camera or a hyperspectral camera. We demonstrated 2D visualization of temperature and oxygen distribution in a model soil system. The RGB camera provided rapid and cost-effective imaging, while the hyperspectral camera offered more detailed spectral information despite some limitations. Our findings revealed the formation of a stable temperature gradient and oxygen depletion, driven by water content and temperature-sensitive microbial activity. This dual O2/T sensor, with further potential improvements, shows considerable promise for advanced multiparameter sensing in complex biological and environmental studies, providing deeper insights into dynamic microenvironments.
化学梯度在生物系统中至关重要,影响着土壤中微生物活动和养分循环等过程。微传感器等传统工具可提供高分辨率数据,但仅限于一维测量。平面光电二极管可进行二维(2D)和三维(3D)化学成像,但通常对温度变化很敏感。本研究提出了一种先进的双发射光学传感器,利用改性铂(II)介-四(3,5-二叔丁基苯基)-四(2-叔丁基-1,4-萘醌)卟啉同时测量温度和氧气。通过对铂(II)萘醌卟啉进行叔丁基修饰,优化了热激活延迟荧光和磷光之间的比例,同时提高了在无极性溶剂和聚合物基质(聚苯乙烯)中的溶解度。这种双功能传感器可通过消费级 RGB 相机或高光谱相机实现双参数化学成像。我们演示了模型土壤系统中温度和氧气分布的二维可视化。RGB 相机能快速、经济地进行成像,而高光谱相机尽管有一些局限性,但能提供更详细的光谱信息。我们的研究结果表明,在含水量和对温度敏感的微生物活动的驱动下,形成了稳定的温度梯度和氧气耗竭。这种 O2/T 双传感器具有进一步改进的潜力,在复杂的生物和环境研究中显示出先进的多参数传感技术的巨大前景,为动态微环境提供了更深入的洞察力。
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引用次数: 0
Activatable Multiplexed 19F NMR Probes for Dynamic Monitoring of Biomarkers Associated with Cellular Senescence 用于动态监测细胞衰老相关生物标记物的可激活多重 19F NMR 探针
Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-08-31 DOI: 10.1021/acsmeasuresciau.4c00045
Jian Wang, Donghui Hong, Jili Li, Linlin Wang, Yuqi Xie, Jun Da, Yanlan Liu
Simultaneous detection of multiple biomarkers is crucial to achieve specific and dynamic analysis of cellular senescence, given its intrinsic high heterogeneity. Current approaches for senescence detection largely rely on fluorescence imaging, but fluorescent probes inevitably suffer from issues including autofluorescence and spectral overlap when being applied for the simultaneous detection of multiple biomarkers. Herein, we report an alternative strategy and design activatable multiplexed senoprobes based on 19F NMR for dynamic monitoring of cellular senescence. Differing from previous approaches, our strategy has two unique advantages. First, this strategy utilizes the changes in the 19F chemical shift as the signal output, which features by its fingerprint and quantifiable characters, thereby significantly enhancing the detection throughput toward biomarkers with minimized spectral overlapping. Second, the background signal is minimized, benefiting from the extremely low abundance of F in biological samples, and the detection accuracy can thus be improved. As a proof of concept, two activatable 19F NMR molecular probes are synthesized that specially respond to two key senescence-associated biomarkers (β-gal and ROS) and have been successfully demonstrated for dynamical and quantitative assessment of the changes of these biomarkers in different cellular models of senescence, without causing obvious cytotoxicity. Owing to the flexible molecular design, this work may offer a useful platform to create diversified 19F NMR senoprobes for deep understanding of cellular senescence across a wide range of aging-related diseases.
鉴于细胞衰老的内在高度异质性,同时检测多种生物标记物对于实现细胞衰老的特异性动态分析至关重要。目前的衰老检测方法主要依赖荧光成像,但荧光探针在用于同时检测多种生物标记物时不可避免地会出现自发荧光和光谱重叠等问题。在此,我们报告了一种替代策略,并基于 19F NMR 设计了可激活的多重衰老探针,用于动态监测细胞衰老。与以往的方法不同,我们的策略有两个独特的优势。首先,该策略利用 19F 化学位移的变化作为信号输出,这种信号输出具有指纹特征和可量化特征,从而大大提高了生物标记物的检测通量,并最大限度地减少了光谱重叠。其次,由于生物样本中 F 的丰度极低,本底信号可降至最低,从而提高检测精度。作为概念验证,我们合成了两种可激活的 19F NMR 分子探针,它们能特别响应两种关键的衰老相关生物标志物(β-gal 和 ROS),并已成功用于动态和定量评估这些生物标志物在不同衰老细胞模型中的变化,且不会引起明显的细胞毒性。由于分子设计灵活,这项工作可能为创建多样化的 19F NMR 衰老探针提供了一个有用的平台,有助于深入了解各种衰老相关疾病的细胞衰老情况。
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引用次数: 0
Activatable Multiplexed 19F NMR Probes for Dynamic Monitoring of Biomarkers Associated with Cellular Senescence 用于动态监测细胞衰老相关生物标记物的可激活多重 19F NMR 探针
IF 4.6 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2024-08-31 DOI: 10.1021/acsmeasuresciau.4c0004510.1021/acsmeasuresciau.4c00045
Jian Wang, Donghui Hong, Jili Li*, Linlin Wang, Yuqi Xie, Jun Da and Yanlan Liu*, 

Simultaneous detection of multiple biomarkers is crucial to achieve specific and dynamic analysis of cellular senescence, given its intrinsic high heterogeneity. Current approaches for senescence detection largely rely on fluorescence imaging, but fluorescent probes inevitably suffer from issues including autofluorescence and spectral overlap when being applied for the simultaneous detection of multiple biomarkers. Herein, we report an alternative strategy and design activatable multiplexed senoprobes based on 19F NMR for dynamic monitoring of cellular senescence. Differing from previous approaches, our strategy has two unique advantages. First, this strategy utilizes the changes in the 19F chemical shift as the signal output, which features by its fingerprint and quantifiable characters, thereby significantly enhancing the detection throughput toward biomarkers with minimized spectral overlapping. Second, the background signal is minimized, benefiting from the extremely low abundance of F in biological samples, and the detection accuracy can thus be improved. As a proof of concept, two activatable 19F NMR molecular probes are synthesized that specially respond to two key senescence-associated biomarkers (β-gal and ROS) and have been successfully demonstrated for dynamical and quantitative assessment of the changes of these biomarkers in different cellular models of senescence, without causing obvious cytotoxicity. Owing to the flexible molecular design, this work may offer a useful platform to create diversified 19F NMR senoprobes for deep understanding of cellular senescence across a wide range of aging-related diseases.

鉴于细胞衰老的内在高度异质性,同时检测多种生物标记物对于实现细胞衰老的特异性动态分析至关重要。目前的衰老检测方法主要依赖荧光成像,但荧光探针在用于同时检测多种生物标记物时不可避免地会出现自发荧光和光谱重叠等问题。在此,我们报告了一种替代策略,并基于 19F NMR 设计了可激活的多重衰老探针,用于动态监测细胞衰老。与以往的方法不同,我们的策略有两个独特的优势。首先,该策略利用 19F 化学位移的变化作为信号输出,这种信号输出具有指纹特征和可量化特征,从而大大提高了生物标记物的检测通量,并最大限度地减少了光谱重叠。其次,由于生物样本中 F 的丰度极低,本底信号可降至最低,从而提高检测精度。作为概念验证,我们合成了两种可激活的 19F NMR 分子探针,它们能特别响应两种关键的衰老相关生物标志物(β-gal 和 ROS),并已成功用于动态和定量评估这些生物标志物在不同衰老细胞模型中的变化,且不会引起明显的细胞毒性。由于分子设计灵活,这项工作可能为创建多样化的 19F NMR 衰老探针提供了一个有用的平台,有助于深入了解各种衰老相关疾病的细胞衰老情况。
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ACS Measurement Science Au
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