Bioimaging

生物成像
  • 文章类型: Journal Article
    从焦化等工业应用中排放的二氧化硫(SO2)及其衍生物,运输和食品加工引起了人们对公众健康和环境质量的极大关注。能够检测SO2衍生物的灵敏度和特异性的探针在其法规中发挥关键作用,并最终减轻其对环境和健康的影响,但是荧光探针可以准确地,快速和现场检测食品和环境系统中的SO2衍生物很少报道。在这里,通过调节我们先前报道的高性能HSO3-荧光探针SL的结构,设计并合成了用于亚硫酸氢盐(HSO3-)比率响应的近红外(NIR)荧光探针(ZTX),理论计算和相关文献报告。电子缺陷C=C键与HSO3-之间的迈克尔加成反应破坏了ZTX的π共轭系统并阻断了其分子内电荷转移(ICT)过程,导致紫红色溶液明显褪色,蓝紫色荧光变为浅蓝色荧光。已经证明了利用ZTX对活体动物中的HSO3-进行荧光成像。使用ZTXvia智能手机对食品样品中HSO3-进行定量分析也已成功实施。同时,基于ZTX的测试条用于通过智能手机定量测定环境水样中的HSO3-。因此,ZTX探针可以提供一种新的方法来理解HSO3-,评估食品安全和监测环境,并且具有广泛的应用前景。
    The emission of venenous sulfur dioxide (SO2) and its derivatives from industrial applications such as coking, transportation and food processing has caused great concern about public health and environmental quality. Probes that enable sensitivity and specificity to detect SO2 derivatives play a crucial role in its regulations and finally mitigating its environmental and health impacts, but fluorescent probes that can accurately, rapidly and on-site detect SO2 derivatives in foodstuffs and environmental systems rarely reported. Herein, a near-infrared (NIR) fluorescent probe (ZTX) for the ratiometric response of bisulfite (HSO3-) was designed and synthesized by regulating the structure of high-performance HSO3- fluorescent probe SL previously reported by us based on structural analyses, theoretical calculations and related literature reports. The Michael addition reaction between the electronic-deficient C=C bond and HSO3- destroys ZTX\'s π-conjugation system and blocks its intramolecular charge transfer (ICT) process, resulting in a significant fading of the fuchsia solution and the bluish-purple fluorescence turned light blue fluorescence. Fluorescent imaging of HSO3- in live animals utilizing ZTX has been demonstrated. The quantitative analysis of HSO3- in food samples using ZTXvia a smartphone has been also successfully implemented. Simultaneously, the ZTX-based test strips were utilized to quantificationally determine HSO3- in environmental water samples by a smartphone. Consequently, probe ZTX could provide a new method to understand the physiopathological roles of HSO3-, evaluate food safety and monitor environment, and is promising for broad applications.
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  • 文章类型: Journal Article
    已有效合成了一种新的基于蒽的探针(E)-N'-(1-(蒽-9-基)亚乙基)-2-羟基苯甲酰肼(AHB),并通过各种光谱方法进行了表征。由于具有螯合增强荧光(CHEF)效应的受限光诱导电子转移(PET)机制,它对Al3离子表现出极其选择性和灵敏的荧光传感,荧光强度大大提高。AHB-Al3+复合物在0.498nM显示出极低的检测极限。绑定机制是基于作业的情节提出的,1HNMR滴定,傅里叶变换红外(FT-IR),高分辨率质谱(HRMS),和密度泛函理论(DFT)研究。化学传感器在存在ctDNA的情况下是可重复使用和可逆的。荧光传感器的实际可用性已由测试条套件建立。Further,AHB对Al3+离子诱导的tau蛋白毒性的治疗潜力已经通过金属螯合疗法在果蝇的阿尔茨海默病(AD)模型的眼中进行了测试。AHB显示出巨大的治疗潜力,在眼部表型中具有53.3%的挽救。果蝇肠组织中AHB与Al3+的体内相互作用研究证实了其在生物环境中的传感效率。详细的比较表包括评估AHB的有效性。
    A new anthracene-based probe (E)-N\'-(1-(anthracen-9-yl)ethylidene)-2-hydroxybenzohydrazide (AHB) has been efficiently synthesized and characterized by various spectroscopic methods. It exhibits extremely selective and sensitive fluorometric sensing of Al3+ ions with a large enhancement in the fluorescent intensity due to the restricted photoinduced electron transfer (PET) mechanism with a chelation-enhanced fluorescence (CHEF) effect. The AHB-Al3+ complex shows a remarkably low limit of detection at 0.498 nM. The binding mechanism has been proposed based on Job\'s plot, 1H NMR titration, Fourier transform infrared (FT-IR), high-resolution mass spectrometry (HRMS), and density functional theory (DFT) studies. The chemosensor is reusable and reversible in the presence of ctDNA. The practical usability of the fluorosensor has been established by a test strip kit. Further, the therapeutic potential of AHB against Al3+ ion-induced tau protein toxicity has been tested in the eye of Alzheimer\'s disease (AD) model of Drosophila via metal chelation therapy. AHB shows great therapeutic potential with 53.3% rescue in the eye phenotype. The in vivo interaction study of AHB with Al3+ in the gut tissue of Drosophila confirms its sensing efficiency in the biological environment. A detailed comparison table included evaluates the effectiveness of AHB.
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  • 文章类型: Journal Article
    预测固体制剂体内性能的体外溶出在制剂优化中极为重要。吸收分数(Fa)已用于基于体外-体内相关性(IVIVC)的思想筛选体外溶出方案,但由于Fa的不准确性而未能提高成功率。IVIVC的本质是体外溶出度和体内溶出度之间的相关性。我们试图通过与使用阿立哌唑(APZ)作为模型药物的体内溶出相似来建立体外溶出方案。通过将聚集引起的猝灭(ACQ)荧光团物理嵌入晶格内部以测量体内溶解来制备杂化APZ晶体(APZ-HC)。该过程没有改变APZ的理化性质和结晶度。荧光团照亮APZ晶体,但在APZ-HC溶解在水性介质中时猝灭,能够实时监控完整的APZ-HC。APZ-HC的荧光猝灭和溶解之间的良好相关性证明了完整APZ晶体的可靠定量。记录APZ-HC处理的大鼠荧光强度的残余百分比随时间的变化,从100%的差异转化为体内溶出度。用Fa验证体内溶出度。通过与体内溶出度相比大于50的相似因子建立APZ的体外溶出度曲线。该研究为建立体外溶出方案提供了新的思路和方法。
    In vitro dissolution that predicts the in vivo performance of solid preparations is extremely important in formulation optimization. Fraction absorbed (Fa) has been used to screen in vitro dissolution protocols based on the idea of in vitro-in vivo correlation (IVIVC) but failed to increase the success rate due to the inaccuracy of the Fa. The essence of IVIVC is the correlation between in vitro dissolution and in vivo dissolution. We tried to establish in vitro dissolution protocol via similarity with in vivo dissolution using aripiprazole (APZ) as a model drug. Hybrid APZ crystals (APZ-HCs) were prepared by physically embedding aggregation-caused quenching (ACQ) fluorophores inside the lattice to measure the in vivo dissolution. The process did not change the physicochemical properties and crystallinity of APZ. The fluorophore illuminated APZ crystals but was quenched upon dissolution of APZ-HCs in aqueous media, enabling monitoring intact APZ-HCs in real-time. The good correlation between fluorescent quenching and dissolution of APZ-HCs justified reliable quantification of intact APZ crystals. The residual percentage of fluorescence intensity in rats treated by APZ-HCs was recorded with time, which was converted to in vivo dissolution by the difference from 100%. The in vivo dissolution was validated with the Fa. The in vitro dissolution profile of APZ was set up via a similarity factor larger than 50 in comparison with the in vivo dissolution. The study provides a novel idea and method to establish in vitro dissolution protocol.
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  • 文章类型: Journal Article
    已经报道了许多关于源自各种来源的单色和多色高荧光碳纳米颗粒(FCNP)及其在生物成像中的潜在应用的研究。在这里,研究了从椰枣叶中脱出的多色生物相容性碳纳米颗粒(CNPs)。通过几种显微镜和光谱技术表征提取的CNP。结果表明,CNP是结晶石墨和亲水性的,尺寸范围为4至20nm。未包覆的CNP显示示例性光致发光(PL)性质。当暴露于紫外线(UV)光时,它们也发出明亮的蓝色。此外,还研究了在CNP存在下的体外细胞摄取和细胞活力。通过细胞代谢活性测定(MTT)孵育24和48小时,评估了具有不同浓度的水性CNP的人结肠癌(HCT-116)和乳腺癌(MCF-7)细胞系的细胞活力。组合结果以产生CNP的剂量-反应曲线并评估其毒性的严重性。对于体外细胞成像,CNP显示具有高细胞活力的足够荧光。在激光扫描共聚焦显微镜下,具有HCT-116和MCF-7细胞系的CNPs显示多色荧光发射,包括蓝色,绿色,在405、458和561nm激发时呈红色,分别。这些结果证明,由于其低细胞毒性,来自椰枣叶的未鞘CNPs可用于多种生物医学应用。足够的荧光,环保自然,廉价的生产。
    Numerous studies have been reported on single- and multicolored highly fluorescent carbon nanoparticles (FCNPs) originating from various sources and their potential applications in bioimaging. Herein, multicolored biocompatible carbon nanoparticles (CNPs) unsheathed from date palm fronds were studied. The extracted CNPs were characterized via several microscopic and spectroscopic techniques. The results revealed that the CNPs were crystalline graphitic and hydrophilic in nature with sizes ranging from 4 to 20 nm. The unsheathed CNPs showed exemplary photoluminescent (PL) properties. They also emitted bright blue colors when exposed to ultraviolet (UV) light. Furthermore, in vitro cellular uptake and cell viability in the presence of CNPs were also investigated. The cell viability of human colon cancer (HCT-116) and breast adenocarcinoma (MCF-7) cell lines with aqueous CNPs at different concentrations was assessed by a cell metabolic activity assay (MTT) for 24 and 48 h incubations. The results were combined to generate dose-response curves for the CNPs and evaluate the severity of their toxicity. The CNPs showed adequate fluorescence with high cell viability for in vitro cell imaging. Under the laser-scanning confocal microscope, the CNPs with HCT-116 and MCF-7 cell lines showed multicolor fluorescence emissions, including blue, green, and red colors when excited at 405, 458, and 561 nm, respectively. These results prove that unsheathed CNPs from date palm fronds can be used in diverse biomedical applications because of their low cytotoxicity, adequate fluorescence, eco-friendly nature, and cheap production.
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  • 文章类型: Journal Article
    Platelets and their subcellular components (e.g., dense granules) are essential components in hemostasis. Understanding their chemical heterogeneities at the sub-micrometer scale, particularly their activation during hemostasis and production of platelet-derived extracellular vesicles, may provide important insights into their mechanisms; however, this has rarely been investigated, mainly owing to the lack of appropriate chemical characterization tools at nanometer scale. Here, the use of scanning transmission X-ray microscopy (STXM) combined with X-ray absorption near edge structure (XANES) to characterize human platelets and their subcellular components at the carbon K-edge and calcium L2,3-edge, is reported. STXM images can identify not only the spatial distribution of subcellular components in human platelets, such as dense granules (DGs) with sizes of ~200 nm, but also their granule-to-granule chemical heterogeneities on the sub-micrometer scale, based on their XANES spectra. The calcium distribution map as well as the principal component analysis of the STXM image stacks clearly identified the numbers and locations of the calcium-rich DGs within human platelets. Deconvolution of the carbon K-edge XANES spectra, extracted from various locations in the platelets, showed that amide carbonyl and carboxylic acid functional groups were mainly found in the cytoplasm, while ketone-phenol-nitrile-imine, aliphatic, and carbonate functional groups were dominant in the platelet DGs. These observations suggest that platelet DGs are most likely composed of calcium polyphosphate associated with adenosine triphosphate (ATP) and adenosine diphosphate (ADP), with significant granule-to-granule variations in their compositions, while the cytoplasm regions of platelets contain significant amounts of proteins.
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  • 文章类型: Journal Article
    The aim of the research was to design an advanced analytical tool for the precise characterization of microbial aggregates from biofilms formed on food-processing surfaces. The approach combined imaging flow cytometry with a machine learning-based interpretation protocol. Biofilm samples were collected from three diagnostic points of the food-processing lines at two independent time points. The samples were investigated for the complexity of microbial aggregates and cellular metabolic activity. Thus, aggregates and singlets of biofilm-associated microbes were simultaneously examined for the percentages of active, mid-active, and nonactive (dead) cells to evaluate the physiology of the microbial cells forming the biofilm structures. The tested diagnostic points demonstrated significant differences in the complexity of microbial aggregates. The significant percentages of the bacterial aggregates were associated with the dominance of active microbial cells, e.g., 75.3% revealed for a mushroom crate. This confirmed the protective role of cellular aggregates for the survival of active microbial cells. Moreover, the approach enabled discriminating small and large aggregates of microbial cells. The developed tool provided more detailed characteristics of bacterial aggregates within a biofilm structure combined with high-throughput screening potential. The designed methodology showed the prospect of facilitating the detection of invasive biofilm forms in the food industry environment.
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  • 文章类型: Journal Article
    开发了具有优异荧光性能的近红外荧光团,一个大的斯托克斯位移,良好的生物相容性已成为近年来荧光成像领域的研究热点。基于量子化学计算和合理的分子设计策略,开发了一种新的近红外荧光团,其特征在于合成简单,易于结构修改,和大的斯托克斯位移(105nm)。此外,使用简单的结构修饰合成了两种新的“可激活”荧光探针QN-Cys和QN-DNP。探针QN-Cys可以高灵敏度(LOD=128nM)和高选择性识别Cys,其荧光强度在5~35μM范围内与Cys浓度呈良好的线性关系。此外,QN-Cys探针能有效区分Cys与Hcy和GSH,并成功应用于人血清中Cys的检测和成像,细胞,还有斑马鱼.探针QN-DNP对苯硫酚表现出良好的特异性和敏感性(LOD=78nM)荧光响应,其荧光强度在5~30μM范围内与苯硫酚浓度呈良好的线性关系。此外,成功应用于实际水样中苯硫酚的检测,回收率(97-102%),在活细胞中成像苯硫酚,斑马鱼和老鼠。值得注意的是,QN-DNP探针可用于观察苯硫酚在小鼠体内的分布。
    The development of a near-infrared fluorophore with excellent fluorescence performance, a large Stokes shift, and good biocompatibility has become a focus in the field of fluorescence imaging in recent years. Based on quantum chemistry calculations and reasonable molecular design strategies, a new NIR fluorophore was developed and characterized by simple synthesis, easy structural modification, and a large Stokes shift (105 nm). Furthermore, two new \"activatable\" fluorescent probes QN-Cys and QN-DNP were synthesized using a simple structural modification. The probe QN-Cys can recognize Cys with high sensitivity (LOD = 128 nM) and high selectivity, and its fluorescence intensity has a good linear relationship with the Cys concentration in the range of 5-35 μM. Furthermore, probe QN-Cys can effectively distinguish Cys from Hcy and GSH, and was successfully applied to the detection and imaging of Cys in human serum, cells, and zebrafish. The probe QN-DNP showed a good specific and sensitive (LOD = 78 nM) fluorescence response to thiophenol, and its fluorescence intensity has a good linear relationship with the thiophenol concentration in the range of 5-30 μM. Furthermore, it was successfully applied to detect thiophenol in real water samples with good recoveries (97-102%), and image thiophenol in living cells, zebrafish and mice. Notebly, the QN-DNP probe could be applied to visualize the distribution of thiophenol in the mice.
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  • 文章类型: Journal Article
    肿瘤血管和血管生成在肿瘤进展中起着至关重要的作用。因此,它们的可视化对社区至关重要。在这项原理证明研究中,我们已经建立了一个新的跨模态成像(CMI)管道来表征完全相同的小鼠肿瘤跨越尺度和穿透深度,使用黑色素瘤癌的原位模型。这允许获取单个肿瘤的一组全面的血管参数。工作流程可视化不同长度尺度的毛细血管,将它们置于整个肿瘤血管网络的背景下,并允许通过不同的模态对血管密度和形态进行量化和比较。工作流程增加了有关缺氧和血液流速的信息。CMI方法包括完善的技术,如磁共振成像(MRI),正电子发射断层扫描(PET),计算机断层扫描(CT),和超声(美国),和最近进入临床前发现的模式,如光学相干断层扫描(OCT)和高分辨率表观显微镜(HREM)。这种新颖的CMI平台建立了使用广泛的图像处理管道组合这些技术的可行性。尽管在跨空间分辨率的微观和宏观数据的整合方面存在挑战,我们还建立了一个开源管道,用于不同多尺度数据集的半自动共同注册,这使得真正相关的血管成像。虽然专注于肿瘤脉管系统,我们的CMI平台可用于解决癌症生物学中的许多研究问题。
    Tumor vasculature and angiogenesis play a crucial role in tumor progression. Their visualization is therefore of utmost importance to the community. In this proof-of-principle study, we have established a novel cross-modality imaging (CMI) pipeline to characterize exactly the same murine tumors across scales and penetration depths, using orthotopic models of melanoma cancer. This allowed the acquisition of a comprehensive set of vascular parameters for a single tumor. The workflow visualizes capillaries at different length scales, puts them into the context of the overall tumor vessel network and allows quantification and comparison of vessel densities and morphologies by different modalities. The workflow adds information about hypoxia and blood flow rates. The CMI approach includes well-established technologies such as magnetic resonance imaging (MRI), positron emission tomography (PET), computed tomography (CT), and ultrasound (US), and modalities that are recent entrants into preclinical discovery such as optical coherence tomography (OCT) and high-resolution episcopic microscopy (HREM). This novel CMI platform establishes the feasibility of combining these technologies using an extensive image processing pipeline. Despite the challenges pertaining to the integration of microscopic and macroscopic data across spatial resolutions, we also established an open-source pipeline for the semi-automated co-registration of the diverse multiscale datasets, which enables truly correlative vascular imaging. Although focused on tumor vasculature, our CMI platform can be used to tackle a multitude of research questions in cancer biology.
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  • 文章类型: Journal Article
    通过简单的纯化步骤,通过一锅Claisen-Schmidt反应合成了具有电子推挽效应的查尔酮系列(3a-f)。这些化合物表现出强烈的发射,在极性溶剂(DMSO,MeOH,和PBS),并显示出良好的光稳定性。因此,3a-f用于细胞成像。孵育3小时后,与正常细胞(HEK-293)相比,癌细胞(HepG2)中的绿色荧光明显更亮,表明在癌细胞中优先积累。此外,所有化合物在24小时内对癌细胞表现出比正常细胞(IC50值>100μM)更高的细胞毒性(IC50值范围为45至100μM)。此外,研究了查耳酮3a-f的抗菌性能。有趣的是,3a-f对大肠杆菌和金黄色葡萄球菌具有抗菌活性,最小杀菌浓度(MBC)为0.10-0.60mg/mL(375-1000µM),表明它们对革兰氏阴性和革兰氏阳性细菌的潜在抗菌活性。因此,该系列查尔酮衍生的荧光染料易于合成,显示出开发抗生素和癌细胞染色剂的巨大潜力。
    A chalcone series (3a-f) with electron push-pull effect was synthesized via a one-pot Claisen-Schmidt reaction with a simple purification step. The compounds exhibited strong emission, peaking around 512-567 nm with mega-stokes shift (∆λ = 93-139 nm) in polar solvents (DMSO, MeOH, and PBS) and showed good photo-stability. Therefore, 3a-f were applied in cellular imaging. After 3 h of incubation, green fluorescence was clearly brighter in cancer cells (HepG2) compared to normal cells (HEK-293), suggesting preferential accumulation in cancer cells. Moreover, all compounds exhibited higher cytotoxicity within 24 h toward cancer cells (IC50 values ranging from 45 to 100 μM) than normal cells (IC50 value >100 μM). Furthermore, the antimicrobial properties of chalcones 3a-f were investigated. Interestingly, 3a-f exhibited antibacterial activities against Escherichia coli and Staphylococcus aureus, with minimum bactericidal concentrations (MBC) of 0.10-0.60 mg/mL (375-1000 µM), suggesting their potential antibacterial activity against both Gram-negative and Gram-positive bacteria. Thus, this series of chalcone-derived fluorescent dyes with facile synthesis shows great potential for the development of antibiotics and cancer cell staining agents.
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  • 文章类型: Journal Article
    我们报告了含有具有聚集诱导发射特性的发光体的两亲性氨基糖苷缀合物作为转染试剂的表征。这些固有的发光转染载体能够通过静电相互作用结合质粒DNA;由于发光体核心的分子内运动的限制,这种结合导致发射“开启”信号。发光的阳离子两亲物有效地将质粒DNA转移到哺乳动物细胞中(HeLa,HEK293T),如红色荧光蛋白标记的表达所证明。通过显微镜以及ζ电位和动态光散射测量研究了聚集体的形态。使用发光阳离子两亲物的转染效率类似于金标准转染试剂Lipofectamine®2000的转染效率。
    We report the characterization of amphiphilic aminoglycoside conjugates containing luminophores with aggregation-induced emission properties as transfection reagents. These inherently luminescent transfection vectors are capable of binding plasmid DNA through electrostatic interactions; this binding results in an emission \"on\" signal due to restriction of intramolecular motion of the luminophore core. The luminescent cationic amphiphiles effectively transferred plasmid DNA into mammalian cells (HeLa, HEK 293T), as proven by expression of a red fluorescent protein marker. The morphologies of the aggregates were investigated by microscopy as well as ζ-potential and dynamic light-scattering measurements. The transfection efficiencies using luminescent cationic amphiphiles were similar to that of the gold-standard transfection reagent Lipofectamine® 2000.
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