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A new twist on PIFE: photoisomerisation-related fluorescence enhancement. PIFE的一个新转折:与光异构化相关的荧光增强。
IF 2.4 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-10-12 DOI: 10.1088/2050-6120/acfb58
Evelyn Ploetz, Benjamin Ambrose, Anders Barth, Richard Börner, Felix Erichson, Achillefs N Kapanidis, Harold D Kim, Marcia Levitus, Timothy M Lohman, Abhishek Mazumder, David S Rueda, Fabio D Steffen, Thorben Cordes, Steven W Magennis, Eitan Lerner

PIFE was first used as an acronym for protein-induced fluorescence enhancement, which refers to the increase in fluorescence observed upon the interaction of a fluorophore, such as a cyanine, with a protein. This fluorescence enhancement is due to changes in the rate ofcis/transphotoisomerisation. It is clear now that this mechanism is generally applicable to interactions with any biomolecule. In this review, we propose that PIFE is thereby renamed according to its fundamental working principle as photoisomerisation-related fluorescence enhancement, keeping the PIFE acronym intact. We discuss the photochemistry of cyanine fluorophores, the mechanism of PIFE, its advantages and limitations, and recent approaches to turning PIFE into a quantitative assay. We provide an overview of its current applications to different biomolecules and discuss potential future uses, including the study of protein-protein interactions, protein-ligand interactions and conformational changes in biomolecules.

PIFE最初被用作蛋白质诱导荧光增强的缩写,指的是在荧光团(如花青)与蛋白质相互作用时观察到的荧光增加。这种荧光增强是由于顺式/反式光异构化速率的变化。现在很清楚,这种机制通常适用于与任何生物分子的相互作用,在这篇综述中,我们建议根据其基本工作原理将PIFE重命名为与光异构化相关的荧光增强,保持PIFE首字母缩写的完整性。我们讨论了菁荧光团的光化学、PIFE的机制、其优点和局限性,以及将PIFE转化为定量分析的最新方法。我们概述了它目前在不同生物分子中的应用,并讨论了未来的潜在用途,包括蛋白质-蛋白质相互作用、蛋白质-配体相互作用和生物分子构象变化的研究。
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引用次数: 0
Comparison of MSD analysis from single particle tracking with MSD from images. Getting the best of both worlds. 来自单粒子跟踪的MSD分析与来自图像的MSD的比较。两全其美。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-10-12 DOI: 10.1088/2050-6120/acfd7e
Constanza Kettmayer, Enrico Gratton, Laura C Estrada

Fluorescence microscopy can provide valuable information about cell interior dynamics. Particularly, mean squared displacement (MSD) analysis is widely used to characterize proteins and sub-cellular structures' mobility providing the laws of molecular diffusion. The MSD curve is traditionally extracted from individual trajectories recorded by single-particle tracking-based techniques. More recently, image correlation methods like iMSD have been shown capable of providing averaged dynamic information directly from images, without the need for isolation and localization of individual particles. iMSD is a powerful technique that has been successfully applied to many different biological problems, over a wide spatial and temporal scales. The aim of this work is to review and compare these two well-established methodologies and their performance in different situations, to give an insight on how to make the most out of their unique characteristics. We show the analysis of the same datasets by the two methods. Regardless of the experimental differences in the input data for MSD or iMSD analysis, our results show that the two approaches can address equivalent questions for free diffusing systems. We focused on studying a range of diffusion coefficients between D = 0.001μm2s-1and D = 0.1μm2s-1, where we verified that the equivalence is maintained even for the case of isolated particles. This opens new opportunities for studying intracellular dynamics using equipment commonly available in any biophysical laboratory.

荧光显微镜可以提供关于细胞内部动力学的有价值的信息。特别地,均方位移(MSD)分析被广泛用于表征蛋白质和亚细胞结构的迁移率,提供分子扩散规律。MSD曲线传统上是从基于单粒子跟踪的技术记录的单个轨迹中提取的。最近,像iMSD这样的图像相关方法已经被证明能够直接从图像中提供平均的动态信息,而不需要分离和定位单个粒子。iMSD是一种强大的技术,已在广泛的空间和时间尺度上成功应用于许多不同的生物学问题。这项工作旨在回顾和比较这两种公认的方法及其在不同情况下的表现,以深入了解如何充分利用它们的独特特征。我们展示了用这两种方法对相同数据集的分析。不管MSD或iMSD分析输入数据的实验差异如何,我们的结果表明,这两种方法可以解决自由扩散系统的等效问题。我们专注于研究D=0.001µm2/s和D=0.1µm2/s之间的扩散系数范围,其中我们验证了即使在孤立粒子的情况下也保持了等效性。这为使用任何生物物理实验室中常见的设备研究细胞内动力学开辟了新的机会。
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引用次数: 0
Emission color tuning and dual-mode luminescence thermometry design in Dy3+/Eu3+co-doped SrMoO4phosphors. Dy3+/Eu3+共掺杂SrMoO4磷的发射颜色调谐和双模发光测温设计。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-10-12 DOI: 10.1088/2050-6120/acf97b
Vaibhav Chauhan, Prashant Dixit, Prashant Kumar Pandey, Satyam Chaturvedi, Praveen Chandra Pandey

The challenge of building a highly reliable contactless temperature probe with high sensitivity, good temperature-induced color discriminability, and economical synthesis has prompted the research community to work in the field of rare-earth-based luminescence thermometry. Moreover, the fast-growing market for optoelectronic devices has increased the demand for tunable color-emitting phosphors. In this study, Dy3+/Eu3+co-doped SrMoO4phosphors were developed as tunable color-emitting source and dual-mode luminescence thermometer. A facile and cost-effective auto-combustion method was used to synthesize the phosphors. Our work demonstrates a viable scheme for tailoring the emission of single-phase phosphors by precisely controlling the dopant concentrations and by modulating excitation wavelength. The overall emission is tuned from greenish-yellow to white and greenish-yellow to reddish-orange. A detailed energy transfer process from the host to the Ln3+ions and between the Ln3+ions is discussed. Further, anti-thermal quenching in the emission of Dy3+ion is observed when excited with 297 nm. The dual-mode luminescence thermometry has been studied by analyzing the fluorescence intensity ratio of Dy3+and Eu3+ions upon excitation at 297 nm. The maximum relative sensitivity value for 4% Eu3+co-doped SrMoO4:4%Dy3+phosphor is 1.46% K-1at 300 K. Furthermore, the configurational coordinate diagram is presented to elucidate the nature of temperature-dependent emission. Therefore, our research opens up new avenues for the development of color-tunable luminescent materials for various optoelectronic and temperature-sensing applications.

构建一种具有高灵敏度、良好的温度诱导颜色辨别性和经济合成的高度可靠的非接触式温度探针的挑战促使研究界致力于稀土发光测温领域。此外,快速增长的光电子器件市场增加了对可调谐彩色发光磷光体的需求。本研究开发了Dy3+/Eu3+共掺杂SrMoO4磷作为可调谐彩色发射源和双模发光温度计。采用一种简便、经济的自燃烧方法合成了荧光粉。我们的工作证明了一种通过精确控制掺杂剂浓度和调制激发波长来定制单相磷光体发射的可行方案。整体发射从绿黄色调为白色,从绿黄色变为红橙色。讨论了从主体到Ln3+离子以及Ln3+阳离子之间的详细能量转移过程。此外,当用297nm激发时,观察到Dy3+离子发射中的抗热猝灭。通过分析Dy3+和Eu3+离子在297nm激发时的荧光强度比,研究了双模发光测温法。4%Eu3+共掺杂SrMoO4:4%Dy3+荧光粉在300K时的最大相对灵敏度值为1.46%K-1。此外,还给出了组态坐标图来阐明温度相关发射的性质。因此,我们的研究为开发用于各种光电和温度传感应用的颜色可调发光材料开辟了新的途径。
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引用次数: 0
Carbon dots derived from citric acid and urea as fluorometric probe for determining melamine contamination in infant formula sample. 柠檬酸和尿素衍生的碳点作为荧光探针,用于测定婴儿配方奶粉样品中的三聚氰胺污染。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-10-12 DOI: 10.1088/2050-6120/acf547
Souliyanh Phimmasone, Pornthip Boonsri, Weena Siangproh, Nuanlaor Ratanawimarnwong, Piyada Jittangprasert, Thitirat Mantim, Nunticha Limchoowong, Kriangsak Songsrirote

Melamine has been intentionally added into food products to increase the protein count at less cost, especially in dairy products for infant resulting in serious adverse effects on health of consumers. Therefore, this study aimed to develop a method to quantify melamine in dairy products based on the change of fluorescent properties of carbon dots (CDs) as sensing probe. CDs with green-fluorescent emission were synthesized from citric acid and urea under microwave irradiation. The synthesized CDs emitted fluorescence at the maximum wavelength of 538 nm with excitation wavelength of 410 nm. Thus, they provided high sensitivity and selectivity on melamine detection by which fluorescent emission of the CDs was increasingly quenched upon increasing melamine concentrations. Optimal conditions for melamine determination using the CDs was under pH 6, volume ratio between CDs and sample of 2:8 and reaction time of 15 min. The developed method provided high precision of melamine determination with less than 5% of %RSD (n = 5), wide detection range from 1.0 to 200.0 ppm, and high sensitivity with limit of detection (LOD) of 0.47 ppm and limit of quantification (LOQ) of 1.56 ppm, which is within the regulated level by the Food and Drug Administration of the United States for melamine in dairy products. Several analytical characterization techniques were conducted to elucidate the reaction mechanism between CDs and melamine, and the hydrogen bonding interaction was proposed.

三聚氰胺被有意添加到食品中,以降低成本增加蛋白质含量,尤其是在婴儿乳制品中,这对消费者的健康造成了严重的不利影响。因此,本研究旨在开发一种基于碳点(CD)作为传感探针的荧光特性变化来定量乳制品中三聚氰胺的方法。以柠檬酸和尿素为原料,在微波辐射下合成了具有绿色荧光发射的CDs。合成的CDs在最大波长538nm处发射荧光,激发波长410nm。因此,它们对三聚氰胺检测提供了高灵敏度和选择性,通过该检测,随着三聚氰胺浓度的增加,CD的荧光发射越来越熄灭。用CDs测定三聚氰胺的最佳条件是pH为6,CDs与样品的体积比为2:8,反应时间为15min。该方法的三聚氰胺测定精度高,RSD小于5%(n=5),检测范围宽,从1.0到200.0ppm,灵敏度高,检测限(LOD)为0.47 ppm,定量限(LOQ)为1.56 ppm,在美国食品药品监督管理局对乳制品中三聚氰胺的监管水平内。采用多种分析表征技术来阐明CDs与三聚氰胺之间的反应机理,并提出了氢键相互作用。
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引用次数: 0
Photo-physical characterization of high triplet yield brominated fluoresceins by transient state (TRAST) spectroscopy. 通过瞬态(TRAST)光谱对高三重产率溴化荧光素的光物理表征。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-10-03 DOI: 10.1088/2050-6120/acfb59
Baris Demirbay, Glib Baryshnikov, Martin Haraldsson, Joachim Piguet, Hans Ågren, Jerker Widengren

Photo-induced dark transient states of fluorophores can pose a problem in fluorescence spectroscopy. However, their typically long lifetimes also make them highly environment sensitive, suggesting fluorophores with prominent dark-state formation yields to be used as microenvironmental sensors in bio-molecular spectroscopy and imaging. In this work, we analyzed the singlet-triplet transitions of fluorescein and three synthesized carboxy-fluorescein derivatives, with one, two or four bromines linked to the anthracence backbone. Using transient state (TRAST) spectroscopy, we found a prominent internal heavy atom (IHA) enhancement of the intersystem crossing (ISC) rates upon bromination, inferred by density functional theory calculations to take place via a higher triplet state, followed by relaxation to the lowest triplet state. A corresponding external heavy atom (EHA) enhancement was found upon adding potassium iodide (KI). Notably, increased KI concentrations still resulted in lowered triplet state buildup in the brominated fluorophores, due to relatively lower enhancements in ISC, than in the triplet decay. Together with an antioxidative effect on the fluorophores, adding KI thus generated a fluorescence enhancement of the brominated fluorophores. By TRAST measurements, analyzing the average fluorescence intensity of fluorescent molecules subject to a systematically varied excitation modulation, dark state transitions within very high triplet yield (>90%) fluorophores can be directly analyzed under biologically relevant conditions. These measurements, not possible by other techniques such as fluorescence correlation spectroscopy, opens for bio-sensing applications based on high triplet yield fluorophores, and for characterization of high triplet yield photodynamic therapy agents, and how they are influenced by IHA and EHA effects.

荧光团的光诱导暗瞬态会在荧光光谱中造成问题。然而,它们通常的长寿命也使它们对环境高度敏感,这表明具有显著暗态形成产率的荧光团可以用作生物分子光谱和成像中的微环境传感器。在这项工作中,我们分析了荧光素和三种合成的羧基荧光素衍生物的单线态-三重态跃迁,其中一个、两个或四个溴与蒽骨架相连。使用瞬态(TRAST)光谱,我们发现溴化时系统间交叉(ISC)速率的内部重原子(IHA)显著增强,密度泛函理论计算推断,这是通过较高的三重态发生的,然后弛豫到最低的三重状态。在加入碘化钾(KI)后,发现了相应的外部重原子(EHA)增强。值得注意的是,KI浓度的增加仍然导致溴化荧光团中三重态积累的降低,这是由于ISC的增强相对低于三重态衰变。与对荧光团的抗氧化作用一起,添加KI从而产生溴化荧光团的荧光增强。通过TRAST测量,分析受到系统变化的激发调制的荧光分子的平均荧光强度,可以在生物相关条件下直接分析非常高的三重态产率(>90%)荧光团内的暗态跃迁。这些测量是荧光相关光谱等其他技术无法实现的,可用于基于高三重产率荧光团的生物传感应用,以及高三重产量光动力治疗剂的表征,以及它们如何受到IHA和EHA效应的影响。
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引用次数: 0
Green synthesis of fluorescent carbon dots from Robinia hispida L. leaves for selective detection of Hg (II). 从Robinia hispida L.叶中绿色合成荧光碳点用于汞的选择性检测(II)。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-09-26 DOI: 10.1088/2050-6120/acf97c
Aysel Başoğlu

In this study, Robinia hispida L leaves (RH) was used as a precursor for the first time to synthesize fluorescent carbon dots (CDs) with stable blue fluorescence by a single-step hydrothermal synthesis method. Notably, the innovative approach eliminates the necessity for toxic chemicals or hazardous substances, marking a significant advancement in the field. The synthesized CDs demonstrate CDs demonstrates the predominance of spherical shapes with an average size of 11.63 ± 1.92 nm. The CDs not only exhibit an enhanced fluorescent efficiency with a relatively high quantum yield of up to 6.8%, but they also possess the potential for direct utilization in the selective determination of Hg(II) through fluorescence quenching, even without any functionalization. Under the optimized conditions at a pH of 7.0, a robust linear correlation was found to exist between the fluorescence intensity and the concentration of Hg (II) within the range of 5-17.5μM, exhibiting a detection limit (3σ) of 1.5μM. Additionally, this methodology was effectively employed to successfully detect Hg (II) ions in various aqueous samples, including tap water, spring water, drinking water, and a certified reference material (CRM-SA-C Sandy Soil C). The spike recoveries of 97.6%-101.6% with less than 2.7% variability were performed on all samples.

本研究首次以大叶Robinia hispida L leaves(RH)为前驱体,采用一步水热合成法合成了具有稳定蓝色荧光的荧光碳点(CDs)。值得注意的是,这一创新方法消除了有毒化学品或危险物质的必要性,标志着该领域取得了重大进展。合成的CDs显示CDs以球形为主,平均尺寸为11.63±1.92nm。CDs不仅表现出增强的荧光效率,具有高达6.8%的相对高的量子产率,而且它们还具有通过荧光猝灭直接用于Hg(II)的选择性测定的潜力,即使没有任何功能化。在pH为7.0的优化条件下,发现荧光强度和Hg(II)浓度在5-17.5μM范围内存在稳健的线性相关性,检测极限(3σ)为1.5μM,饮用水和经认证的参考材料(CRM-SA-C Sandy Soil C)。所有样品的加标回收率为97.6%-101.6%,变异性小于2.7%。
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引用次数: 0
Measuring protein-membrane interaction through radial fluorescence correlation in 2 dimensions. 通过二维径向荧光相关测量蛋白质-膜相互作用。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-09-18 DOI: 10.1088/2050-6120/acf118
Natalia Philipp, Enrico Gratton, Laura Estrada

The cell membrane has a fundamental role in the cell life cycle but there's still much to be learned about its heterogeneous structure, regulation, and protein interaction. Additionally, the protein-membrane interaction is often overlooked when studying specific protein dynamics. In this work, we present a new tool for a better understanding of protein dynamics and membrane function using live cells and fast non-invasive techniques without the need for individual particle tracking. To this end, we used the 2D-pair correlation function (2D-pCF) to study protein interactions across cellular membranes. We performed numerical simulations and confocal experiments using a GAP-mEGFP fusion construct known to interact with the plasmatic membrane. Our results demonstrate that based on a quantitative correlation analysis as the 2D pair correlation of the signal intensities, is possible to characterize protein-membrane interactions in live systems and real-time. Combining experimental and numerical results this work presents a new powerful approach to the study of the dynamic protein-membrane interaction.

细胞膜在细胞生命周期中起着至关重要的作用,但关于其异质结构、调控和蛋白质相互作用还有很多需要了解的。此外,在研究特定的蛋白质动力学时,蛋白质-膜相互作用往往被忽视。在这项工作中,我们提出了一种新的工具,可以更好地了解蛋白质动力学和膜功能,使用活细胞和快速非侵入性技术,而无需单个颗粒跟踪。为此,我们使用2D-pair相关函数(2D-pCF)来研究细胞膜上的蛋白质相互作用。我们使用已知与质膜相互作用的GAP-mEGFP融合结构进行了数值模拟和共聚焦实验。我们的研究结果表明,基于定量的相关分析,如信号强度的二维对相关,可以实时表征蛋白质-膜相互作用。结合实验和数值结果,本工作为研究蛋白质-膜的动态相互作用提供了一种新的有力方法。
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引用次数: 0
Multimodal fluorescence imaging and spectroscopic techniques for oral cancer screening: a real-time approach. 口腔癌筛查的多模态荧光成像和光谱技术:一种实时方法。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-09-13 DOI: 10.1088/2050-6120/acf6ac
Pramila Thapa, Veena Singh, Sunil Bhatt, Kiran Maurya, Virendra Kumar, Vivek Nayyar, Kiran Jot, Deepika Mishra, Anurag Shrivastava, Dalip Singh Mehta

The survival rate of oral squamous cell carcinoma (OSCC) patients is very poor, but it can be improved using highly sensitive, specific, and accurate techniques. Autofluorescence and fluorescence techniques are very sensitive and helpful in cancer screening; being directly linked with the molecular levels of human tissue, they can be used as a quantitative tool for cancer detection. Here, we report the development of multi-modal autofluorescence and fluorescence imaging and spectroscopic (MAF-IS) smartphone-based systems for fast and real-time oral cancer screening. MAF-IS system is indigenously developed and offers the advantages of being a low-cost, handy, non-contact, non-invasive, and easily operable device that can be employed in hospitals, including low-resource settings. In this study, we report the results of 43 individuals with 28 OSCC and 15 oral potentially malignant disorders (OPMDs), i.e., epithelial dysplasia and oral submucous fibrosis, using the developed devices. We observed a red shift in fluorescence emission spectrain vivo. We found red-shift of 7.72 ± 6 nm, 3 ± 4.36 nm, and 1.33 ± 0.47 nm in the case of OSCC, epithelial dysplasia, and oral submucous fibrosis, respectively, compared to normal. The results were compared with histopathology and found to be consistent. Further, the MAF-IS system provides results in real-time with higher accuracy and sensitivity compared to devices using a single modality. Our system can achieve an accuracy of 97% with sensitivity and specificity of 100% and 94.7%, respectively, even with a smaller number of patients (28 patients of OSCC). The proposed MAF-IS device has great potential for fast screening and diagnosis of oral cancer in the future.

口腔鳞状细胞癌(OSCC)患者的生存率很低,但使用高灵敏度、特异性和准确性的技术可以提高生存率。自体荧光和荧光技术非常敏感,有助于癌症筛查;由于与人体组织的分子水平直接相关,它们可以用作癌症检测的定量工具。在这里,我们报告了基于智能手机的多模态自体荧光和荧光成像和光谱(MAF-IS)系统的发展,用于快速和实时口腔癌筛查。MAF-IS系统是自主开发的,具有低成本、方便、非接触、非侵入性和易于操作的优点,可用于医院,包括资源匮乏的环境。在这项研究中,我们报告了43例患有28例OSCC和15例口腔潜在恶性疾病(OPMDs)(即上皮发育不良和口腔黏膜下纤维化)的患者使用该装置的结果。我们在体内观察到荧光发射光谱的红移。我们发现,与正常人相比,OSCC、上皮发育不良和口腔黏膜下纤维化的红移分别为7.72±6 nm、3±4.36 nm和1.33±0.47 nm。结果与组织病理学比较,发现是一致的。此外,与使用单一模态的设备相比,MAF-IS系统提供的实时结果具有更高的精度和灵敏度。即使在较少的患者(28例OSCC)中,我们的系统也能达到97%的准确率,100%的灵敏度和94.7%的特异性。本文提出的MAF-IS装置在口腔癌的快速筛查和诊断方面具有很大的潜力。
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引用次数: 0
Quasi-equilibrium state based quantification of biological macromolecules in single-molecule localization microscopy. 单分子定位显微镜中基于准平衡态的生物大分子定量。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-09-08 DOI: 10.1088/2050-6120/acf546
Xuecheng Chen, Yaqian Li, Xiaowei Li, Jielin Sun, Daniel M Czajkowsky, Zhifeng Shao

The stoichiometry of molecular components within supramolecular biological complexes is often an important property to understand their biological functioning, particularly within their native environment. While there are well established methods to determine stoichiometryin vitro, it is presently challenging to precisely quantify this propertyin vivo,especially with single molecule resolution that is needed for the characterization stoichiometry heterogeneity. Previous work has shown that optical microscopy can provide some information to this end, but it can be challenging to obtain highly precise measurements at higher densities of fluorophores. Here we provide a simple approach using already established procedures in single-molecule localization microscopy (SMLM) to enable precise quantification of stoichiometry within individual complexes regardless of the density of fluorophores. We show that by focusing on the number of fluorophore detections accumulated during the quasi equilibrium-state of this process, this method yields a 50-fold improvement in precision over values obtained from images with higher densities of active fluorophores. Further, we show that our method yields more correct estimates of stoichiometry with nuclear pore complexes and is easily adaptable to quantify the DNA content with nanodomains of chromatin within individual chromosomes inside cells. Thus, we envision that this straightforward method may become a common approach by which SMLM can be routinely employed for the accurate quantification of subunit stoichiometry within individual complexes within cells.

超分子生物复合物中分子组分的化学计量学通常是了解其生物功能的重要性质,特别是在其天然环境中。虽然有很好的方法来确定体外化学计量学,但目前在体内精确量化这种特性是具有挑战性的,特别是在表征化学计量学异质性所需的单分子分辨率下。以前的工作已经表明,光学显微镜可以为此提供一些信息,但是在更高密度的荧光团下获得高度精确的测量可能具有挑战性。在这里,我们提供了一种简单的方法,使用单分子定位显微镜(SMLM)中已经建立的程序,无论荧光团的密度如何,都可以精确定量单个复合物内的化学计量学。我们表明,通过关注在该过程的准平衡状态中积累的荧光团检测的数量,该方法产生的精度比从具有较高活性荧光团密度的图像中获得的值提高50倍。此外,我们表明,我们的方法对核孔复合物的化学计量学产生了更正确的估计,并且很容易适用于定量细胞内单个染色体内染色质纳米结构域的DNA含量。因此,我们设想这种简单的方法可能成为一种常见的方法,通过这种方法,SMLM可以常规地用于精确定量细胞内单个复合物内的亚基化学计量。
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引用次数: 0
Rapid microwave synthesis of N and S dual-doped carbon quantum dots for natamycin determination based on fluorescence switch-off assay. 荧光关闭法快速微波合成N、S双掺杂碳量子点测定纳他霉素。
IF 3.2 3区 化学 Q3 CHEMISTRY, ANALYTICAL Pub Date : 2023-08-25 DOI: 10.1088/2050-6120/acf119
Ali Abdel-Hakim, Fatallah Belal, Mohamed A Hammad, Mahmoud Hamed Elmaghrabey

Green, one-pot, quick, and easily synthesized nitrogen and sulfur co-doped carbon quantum dots (N,S-CDs) were obtained from cheap and readily available chemicals (sucrose, urea, and thiourea) using a microwave-assisted approach in about 4 min and utilized as a turn-off fluorescent sensor for estimation of natamycin (NAT). First, the effect of N and S doping on the microwave-synthesized CDs' quantum yield was carefully studied. CDs derived from sucrose alone failed to produce a high quantum yield; then, to increase the quantum yield, doping with heteroatoms was carried out using either urea or thiourea. A slight increase in quantum yield was observed upon using thiourea with sucrose, while an obvious enhancement of quantum yield was obtained when urea was used instead of thiourea. Surprisingly, using a combination of urea and thiourea together results in N,S-CDs with the highest quantum yield (53.5%), uniform and small particle size distribution, and extended stability. The fluorescent signal of N,S-CDs was quenched upon addition of NAT due to inner filter effect and static quenching in a manner that allowed for quantitative determination of NAT over a range of 0.5-10.0μg ml-1(LOD = 0.10μg ml-1). The N,S-CDs were applicable for determination of NAT in aqueous humor, eye drops, different environmental water samples, and bread with excellent performance. The selectivity study indicated excellent selectivity of the prepared N,S-CDs toward NAT with little interference from possibly interfering substances. In-silico toxicological evaluation of NAT was conducted to estimate its long-term toxicity and drug-drug interactions. Finally, the preparation of N,S-CDs, and analytical procedure compliance with the green chemistry principles were confirmed by two greenness assessment tools.

绿色,一锅,快速,容易合成氮和硫共掺杂碳量子点(N,S-CDs)从廉价和容易获得的化学品(蔗糖,尿素和硫脲)使用微波辅助方法在大约4分钟内获得,并用作关闭荧光传感器用于估计纳他霉素(NAT)。首先,研究了N和S掺杂对微波合成CDs量子产率的影响。仅从蔗糖中提取的CDs不能产生高量子产率;然后,为了提高量子产率,用尿素或硫脲掺杂杂原子。硫脲与蔗糖一起使用时,量子产率略有提高,而用尿素代替硫脲时,量子产率有明显提高。令人惊讶的是,使用尿素和硫脲的组合得到的N,S-CDs具有最高的量子产率(53.5%),均匀而小的粒径分布和扩展的稳定性。N,S-CDs的荧光信号在加入NAT后由于内部过滤和静态猝灭作用而猝灭,可以在0.5 ~ 10.0μg ml-1(LOD = 0.10μg ml-1)范围内定量测定NAT。N、S-CDs可用于体液、滴眼液、不同环境水样、面包中NAT的测定,性能优良。选择性研究表明,所制备的N,S-CDs对NAT具有良好的选择性,且不受可能干扰物质的干扰。对NAT进行了计算机毒理学评价,以评估其长期毒性和药物-药物相互作用。最后,通过两种绿色评价工具验证了N、S-CDs的制备和分析过程是否符合绿色化学原则。
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Methods and Applications in Fluorescence
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