Surface plasmon resonance imaging

  • 文章类型: Journal Article
    证明了基于表面等离子体共振成像(SPRI)的生物传感器可同时检测过氧化氢(H2O2)和葡萄糖。待检测的H2O2充当氧化剂并蚀刻银膜。该过程逐渐影响共振条件,并因此影响固定角度的反射光强度。银膜的蚀刻速率与H2O2浓度有明确的关系。因此,监测反射光强度在几分钟内逐渐变化,能够准确检测0至200μM(在0.25-50μM的生理范围内)的H2O2浓度,具有低至40nM的显著检测限(LOD)。在这方面,通过Winspall模拟软件预测了响应于银膜厚度减少的表面等离子体共振(SPR)下降的行为。仿真结果与实验结果吻合较好。此外,所提出的方法可用于确定0至10mM的葡萄糖浓度,涵盖3-8mM的生理范围。这是通过观察通过葡萄糖和葡萄糖氧化酶(Gox)之间的酶促氧化反应产生的H2O2来实现的。该传感器具有卓越的灵敏度和选择性,葡萄糖浓度的检测限低至175μM。此外,在一个实际的人血清样品的葡萄糖浓度的精确测量是可实现的建议的传感器,使用标准添加方法。建议的葡萄糖传感器显示出用于常规葡萄糖测试的有希望的前景。雇佣一个没有标签的人,实时,和多重检测方法。©2017ElsevierInc.保留所有权利。
    A surface plasmon resonance imaging (SPRI)-based biosensor is demonstrated for the detection of both hydrogen peroxide (H2O2) and glucose. The H2O2 to be detected acts as an oxidant and etch the silver film. This process gradually effects on resonance condition and consequently the reflected light intensity at a fixed angle. The etching rate of the silver film shows a clear relation with the H2O2 concentration. Therefore, monitoring the reflected light intensity progressively changing over a few minutes, enables accurate detection of H2O2 concentrations ranging from 0 to 200 μM (within physiological range of 0.25-50 μM), with a remarkable limit of detection (LOD) as low as 40 nM. In this regard, the behavior of the surface plasmon resonance (SPR) dip in response to the reduction of the silver film thickness is predicted by Winspall simulation software. These simulation results are in good agreement with the experimental results. Moreover, the proposed method can be applied to determine glucose concentrations ranging from 0 to 10 mM, encompassing the physiological range of 3-8 mM. This is achieved by observing the generated H2O2 through the enzymatic oxidation reaction between glucose and glucose oxidase (Gox). The sensor demonstrates remarkable sensitivity and selectivity, with a detection limit as low as 175 μM for glucose concentration. Furthermore, accurate measurement of glucose concentration in an actual human serum sample is achievable with the proposed sensor, using the standard addition method. The suggested glucose sensor shows promising prospects for use in routine glucose testing, employing a label-free, real-time, and multiplex detection approach.© 2017 Elsevier Inc. All rights reserved.
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  • 文章类型: Journal Article
    糖尿病是一种以高血糖为特征的代谢性疾病,可以通过抑制α-葡萄糖苷酶(α-Glu)和α-淀粉酶(α-Amy)来抵消,负责碳水化合物水解的酶。近几十年来,已经研究了许多天然化合物及其生物启发类似物作为α-Glu和α-Amy抑制剂。然而,尚未有研究致力于评估新木脂聚糖对α-Glu和α-Amy的抑制作用(1)。在这项工作中,我们报告了1的合成和新类似物库。这些化合物的合成是通过以下方法实现的:苯酚烯丙基化,Claisen/应对重新安排,甲基化,Ullmann耦合,去甲基化,苯酚氧化和迈克尔型加成。评估了Obovatol(1)和十种类似物对α-Glu和α-Amy的体外抑制活性。我们的研究强调,天然存在的1种和4种新木脂聚糖类似物(11、22、26和27)比降血糖药物阿卡波糖(α-Amy:34.6µM;α-Glu:248.3µM)更有效,IC50值为6.2-23.6µM的α-Amy和39.8-124.6µM的α-Glu。对接调查验证了抑制结果,强调合成的新木霉聚糖和两种酶之间的最佳相容性。同时圆二色性光谱法检测到α-Glu与所研究的新木脂素相互作用引起的构象变化。通过荧光测量和α-Glu和α-Amy抑制的动力学的详细研究还表明,1、11、22、26和27对α-Glu具有最大的亲和力,而1、11和27对α-Amy具有最大的亲和力。表面等离子体共振成像(SPRI)测量证实,在所研究的化合物中,Neolignan27对这两种酶都有更大的亲和力,从而证实了通过动力学和荧光猝灭获得的结果。最后,在人结肠癌细胞系(HCT-116)上测试所研究化合物的体外细胞毒性。所有这些结果表明,这些基于obovatol的Neolignan类似物在开发新型降血糖药物方面构成了有希望的候选人。
    Diabetes mellitus is a metabolic disease characterized by hyperglycemia, which can be counteracted by the inhibition of α-glucosidase (α-Glu) and α-amylase (α-Amy), enzymes responsible for the hydrolysis of carbohydrates. In recent decades, many natural compounds and their bioinspired analogues have been studied as α-Glu and α-Amy inhibitors. However, no studies have been devoted to the evaluation of α-Glu and α-Amy inhibition by the neolignan obovatol (1). In this work, we report the synthesis of 1 and a library of new analogues. The synthesis of these compounds was achieved by implementing methodologies based on: phenol allylation, Claisen/Cope rearrangements, methylation, Ullmann coupling, demethylation, phenol oxidation and Michael-type addition. Obovatol (1) and ten analogues were evaluated for their in vitro inhibitory activity towards α-Glu and α-Amy. Our investigation highlighted that the naturally occurring 1 and four neolignan analogues (11, 22, 26 and 27) were more effective inhibitors than the hypoglycemic drug acarbose (α-Amy: 34.6 µM; α-Glu: 248.3 µM) with IC5O value of 6.2-23.6 µM toward α-Amy and 39.8-124.6 µM toward α-Glu. Docking investigations validated the inhibition outcomes, highlighting optimal compatibility between synthesized neolignans and both the enzymes. Concurrently circular dichroism spectroscopy detected the conformational changes in α-Glu induced by its interaction with the studied neolignans. Detailed studies through fluorescence measurements and kinetics of α-Glu and α-Amy inhibition also indicated that 1, 11, 22, 26 and 27 have the greatest affinity for α-Glu and 1, 11 and 27 for α-Amy. Surface plasmon resonance imaging (SPRI) measurements confirmed that among the compounds studied, the neolignan 27 has the greater affinity for both enzymes, thus corroborating the results obtained by kinetics and fluorescence quenching. Finally, in vitro cytotoxicity of the investigated compounds was tested on human colon cancer cell line (HCT-116). All these results demonstrate that these obovatol-based neolignan analogues constitute promising candidates in the pursuit of developing novel hypoglycemic drugs.
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  • 文章类型: Journal Article
    表面等离子体共振(SPR)是定量确定分子相互作用的有力工具。SPR成像(SPRi)进一步提高了SPR技术的吞吐量,并提供了详细观察分子相互作用动力学的空间分辨能力。SPRi在生物和化学传感和成像方面愈来愈普遍。然而,由于不完美的光学组件和沿着表面传播的表面等离子体波的离域特征,SPRi遭受低空间分辨率。已经开发了各种各样的方法来提高SPRi的空间分辨率,极大地推动了该方法的发展,并进一步扩展了其可能的应用。在这篇小型评论中,我们介绍了建立高空间分辨率SPRi系统的机制,并介绍了其从棱镜耦合SPRi和SPR显微镜(SPRM)到表面等离子体散射显微镜(SPSM)的实验方案;总结了其令人兴奋的应用,包括分子相互作用分析,分子成像和分析,跟踪单个实体,和单细胞分析;并讨论其近十年来的挑战以及充满希望的未来。
    Surface plasmon resonance (SPR) is a powerful tool for determining molecular interactions quantitatively. SPR imaging (SPRi) further improves the throughput of SPR technology and provides the spatially resolved capability for observing the molecular interaction dynamics in detail. SPRi is becoming more and more popular in biological and chemical sensing and imaging. However, SPRi suffers from low spatial resolution due to the imperfect optical components and delocalized features of propagating surface plasmonic waves along the surface. Diverse kinds of approaches have been developed to improve the spatial resolution of SPRi, which have enormously impelled the development of the methodology and further extended its possible applications. In this minireview, we introduce the mechanisms for building a high-spatial-resolution SPRi system and present its experimental schemes from prism-coupled SPRi and SPR microscopy (SPRM) to surface plasmonic scattering microscopy (SPSM); summarize its exciting applications, including molecular interaction analysis, molecular imaging and profiling, tracking of single entities, and analysis of single cells; and discuss its challenges in recent decade as well as the promising future.
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  • 文章类型: Journal Article
    已开发出一种基于“表面等离子体共振成像(SPRi)”检测技术的新型生物传感器,用于定量“成纤维细胞生长因子23(FGF23)”。FGF23主要在骨组织中作为磷酸性激素产生,分泌时与“成纤维细胞生长因子受体1(FGFR1)”和αKlotho形成三聚体复合物。FGF23刺激磷酸盐排泄并抑制肾脏中活性维生素D的形成。FGF23已被证明在骨癌的发生和转移中起作用。新开发的方法,基于阵列SPRi生物传感器,经过验证——精度,准确度,和选择性是可以接受的,并产生小于±10%的回收率。生物传感器的直线响应范围为1至75pg/mL。检出限为0.033pg/mL,定量限为0.107pg/mL。生物传感器用于确定健康受试者和患有“透明细胞”肾细胞癌(ccRCC)的患者血浆中的FGF23浓度。将获得的结果与通过“酶联免疫吸附测定(ELISA)”测得的结果进行比较。确定的Pearson相关系数为0.994和0.989,表明新开发的生物传感器可用作ELISA的竞争性方法。
    A new biosensor based on the \"surface plasmon resonance imaging (SPRi)\" detection technique for the quantification of \"fibroblast growth factor 23 (FGF23)\" has been developed. FGF23 is mainly produced in bone tissues as a phosphaturic hormone that forms a trimeric complex with \"fibroblast growth factor receptor 1 (FGFR1)\" and αKlotho upon secretion. FGF23 stimulates phosphate excretion and inhibits the formation of active vitamin D in the kidneys. FGF23 has been shown to play a role in bone carcinogenesis and metastasis. The newly developed method, based on the array SPRi biosensor, was validated-the precision, accuracy, and selectivity were acceptable, and yielded less than ±10% recovery. The rectilinear response of the biosensor ranges from 1 to 75 pg/mL. The limit of detection was 0.033 pg/mL, and the limit of quantification was 0.107 pg/mL. The biosensor was used to determine FGF23 concentrations in the blood plasma of healthy subjects and patients with \"clear cell\" renal cell carcinoma (ccRCC). The obtained results were compared with those measured through an \"enzyme-linked immunosorbent assay (ELISA)\". The determined Pearson correlation coefficients were 0.994 and 0.989, demonstrating that the newly developed biosensor can be used as a competitive method for the ELISA.
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  • 文章类型: Journal Article
    低聚合比率是树脂基复合材料中的主要问题。在本文中,金覆盖的二氧化硅纳米粒子对双酚A二甲基丙烯酸缩水甘油酯(Bis-GMA)的物理化学和机械性能的等离子体效应,在1.4mW/cm2的强度下研究三甘醇二甲基丙烯酸酯(TEGDMA)和氨基甲酸酯二甲基丙烯酸酯(UDMA)绿光可光聚合牙科树脂40秒。透射电子显微镜(TEM)显示约350nm的二氧化硅被12-15nm的金纳米颗粒(AuNP)覆盖,标称覆盖率为100%。使用了五种不同浓度的裸露和片状二氧化硅颗粒;在后一种复合材料中,计算的Au重量%为0.0052重量%,0.0104wt%,0.0208wt%,0.04160wt%,和0.0823重量%。片状二氧化硅填充纳米复合材料的等离子体峰与Irgacure784光引发剂的吸收和绿色LED发光峰重叠。使用径向拉伸强度(DTS)分析了使用绿光照明实现的等离子体激元增强聚合的效果,差示扫描量热法(DSC),表面等离子体共振成像(SPRi),和基于拉曼光谱的转化度(DC)。在所有测量数据中,发现具有0.0208重量%的AuNP的值是最大的。根据我们的结果,可以得出结论,片状二氧化硅颗粒在牙科树脂中的应用可以提高复合材料的聚合比和力学参数。
    A low ratio of polymerization is a major problem in resin-based composites. In this paper, the plasmonic effect of gold-covered silica nanoparticles on the physicochemical and mechanical properties of bisphenol A diglycidyl dimethacrylate (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA) and urethane dimethacrylate (UDMA) green light-photopolymerizable dental resin was investigated at an intensity of 1.4 mW/cm2 for 40 s. Transmission electron microscopy (TEM) showed silica of about 350 nm covered with 12-15 nm gold nanoparticles (Au NPs) at 100% nominal coverage. Five different concentrations of bare and patchy silica particles were used; in the latter composite, the calculated Au wt% were 0.0052 wt%, 0.0104 wt%, 0.0208 wt%, 0.04160 wt%, and 0.0823 wt%. The plasmon peak of patchy silica-filled nanocomposite overlapped with the absorption of Irgacure 784 photoinitiator and green LED light emission peak. The effect of plasmon-enhanced polymerization achieved with green light illumination was analyzed using diametral tensile strength (DTS), differential scanning calorimetry (DSC), surface plasmon resonance imaging (SPRi), and degree of conversion (DC) based on Raman spectroscopy. The values of the Au NP with 0.0208 wt% was found to be maximum in all the measured data. Based on our result, it can be concluded that the application of patchy silica particles in dental resin can improve the polymerization ratio and the mechanical parameters of the composite.
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  • 文章类型: Journal Article
    在过程分析或环境监测中,实时记录复杂样品的成分在很长一段时间内提出了巨大的挑战。有希望的解决方案是无标记的技术,例如表面等离子体共振(SPR)光谱法。他们是,然而,通常由于分析物结合的可逆性差而受到限制。在这项工作中,我们介绍了SPR成像与半选择性功能表面和智能数据分析相结合如何识别小分子和化学相似分子。我们的传感器使用由不同比例的氧化石墨烯和还原氧化石墨烯制成的单个功能点,由于不同的结合亲和力,根据分析物产生独特的信号模式。这些模式允许在使用浓度低至50μM的卷积神经网络(CNN)进行分类后区分四个嘌呤碱基。验证和测试集分类精度在使用标准CNN的多个传感器上的多个测量中是恒定的。这有望成为在复杂混合物中开发在线传感器的未来方法。
    In process analytics or environmental monitoring, the real-time recording of the composition of complex samples over a long period of time presents a great challenge. Promising solutions are label-free techniques such as surface plasmon resonance (SPR) spectroscopy. They are, however, often limited due to poor reversibility of analyte binding. In this work, we introduce how SPR imaging in combination with a semi-selective functional surface and smart data analysis can identify small and chemically similar molecules. Our sensor uses individual functional spots made from different ratios of graphene oxide and reduced graphene oxide, which generate a unique signal pattern depending on the analyte due to different binding affinities. These patterns allow four purine bases to be distinguished after classification using a convolutional neural network (CNN) at concentrations as low as 50 μM. The validation and test set classification accuracies were constant across multiple measurements on multiple sensors using a standard CNN, which promises to serve as a future method for developing online sensors in complex mixtures.
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  • 文章类型: Journal Article
    炎症,缺血性卒中患者在事件发生后立即激活的修复和再生机制在对损伤的反应中合作,即使在事件发生几周后,在功能恢复和脑重塑中,也可以通过康复治疗来维持。尽管如此,患者对治疗的反应很难预测,因为缺乏特定的可测量的恢复标志物,这可能是临床量表在患者评估中的补充。考虑到细胞外囊泡(EV)是参与中风损伤反应的多种分子的载体,在本研究中,我们已经确定了一组EV相关分子,这些分子(i)证实了EV在缺血性卒中后的过程中的关键参与,(ii)可能在康复计划开始时描述缺血性中风患者,(iii)可用于预测患者对治疗的反应。通过多路复用表面等离子体共振成像生物传感器,亚急性缺血性卒中患者被证明在血液中的小EV表面上血管内皮生长因子受体2(VEGFR2)和转运蛋白(TSPO)的表达增加。此外,小胶质细胞EV和内皮EV被证明显著参与缺血性卒中后10天以上发生的细胞间通讯,因此成为缺血性卒中后亚急性期患者特征分析和预测其康复的潜在工具.
    The inflammatory, reparative and regenerative mechanisms activated in ischemic stroke patients immediately after the event cooperate in the response to injury, in the restoration of functions and in brain remodeling even weeks after the event and can be sustained by the rehabilitation treatment. Nonetheless, patients\' response to treatments is difficult to predict because of the lack of specific measurable markers of recovery, which could be complementary to clinical scales in the evaluation of patients. Considering that Extracellular Vesicles (EVs) are carriers of multiple molecules involved in the response to stroke injury, in the present study, we have identified a panel of EV-associated molecules that (i) confirm the crucial involvement of EVs in the processes that follow ischemic stroke, (ii) could possibly profile ischemic stroke patients at the beginning of the rehabilitation program, (iii) could be used in predicting patients\' response to treatment. By means of a multiplexing Surface Plasmon Resonance imaging biosensor, subacute ischemic stroke patients were proven to have increased expression of vascular endothelial growth factor receptor 2 (VEGFR2) and translocator protein (TSPO) on the surface of small EVs in blood. Besides, microglia EVs and endothelial EVs were shown to be significantly involved in the intercellular communications that occur more than 10 days after ischemic stroke, thus being potential tools for the profiling of patients in the subacute phase after ischemic stroke and in the prediction of their recovery.
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  • 文章类型: Journal Article
    Phase interrogation surface plasmon resonance (P-SPR) biosensors have the highest sensitivity among different types of surface plasmon resonance (SPR) biosensors. However, P-SPR sensors have small dynamic detection range and complex device configuration. To solve these two problems, we designed a multi-channel P-SPR imaging (mcP-SPRi) sensing platform based on a common-path ellipsometry scheme. A wavelength sequential selection (WSS) technique for P-SPRi sensing is developed to select the optimal sensing wavelengths according to different refractive indexes (RIs) of the samples, so the inconsistency of SPR signal response for different biomolecule types caused by the small dynamic detection range is eliminated. And a dynamic detection range of 3.7×10-3 RIU is achieved, which is the largest among the current mcP-SPRi biosensors. Remarkably, the individual SPR phase image acquisition time has been greatly reduced to 1s by using WSS method instead of whole spectrum scanning, which enables the high-throughput mcP-SPRi sensing.
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  • 文章类型: Journal Article
    无论纳米粒子(NPs)在生物医学应用中的应用前景如何,一些毒性作用增加了人们对这些纳米材料安全性的担忧。尽管NPs毒性的途径是多种多样的,并且取决于许多参数,例如纳米粒子的性质和生化环境,许多研究提供证据表明,NP与生物分子或细胞膜之间的直接接触会导致细胞失活或损伤,并且可能是细胞毒性的主要机制。在这样的背景下,这项工作的重点是开发一种快速准确的方法来表征NP之间的相互作用,通过表面等离子体共振成像(SPRi)技术研究蛋白质和脂质膜。通过监测在SPRi生物芯片上制备的脂质膜上连续注射几次金NP后反射率的变化来评估金NP与模拟膜的相互作用。关于吸附在其上的金NP的总表面浓度密度,比较了具有不同脂质组成的膜上的相互作用。然后,分析了金和银NP与血液蛋白的缔合/解离和解离常数(koff)的动力学曲线。由1-棕榈酰-2-油酰基-甘油-3-磷酸胆碱和胆固醇(POPC/胆固醇)组成的膜上的表面浓度密度比仅在POPC上注射金NP后发现的值高2.5倍二甲基十八烷基铵(POPC/DDAB)。关于蛋白质,金NP显示与纤维蛋白原的优先结合,导致反射率变化的值比其他蛋白质的值高8倍。不同的是,银NP在所有测试蛋白质上显示相似的相互作用,但在免疫球蛋白G(IgG)上的反射率变化比其他测试蛋白质的反射率高2倍。
    Regardless of the promising use of nanoparticles (NPs) in biomedical applications, several toxic effects have increased the concerns about the safety of these nanomaterials. Although the pathways for NPs toxicity are diverse and dependent upon many parameters such as the nature of the nanoparticle and the biochemical environment, numerous studies have provided evidence that direct contact between NPs and biomolecules or cell membranes leads to cell inactivation or damage and may be a primary mechanism for cytotoxicity. In such a context, this work focused on developing a fast and accurate method to characterize the interaction between NPs, proteins and lipidic membranes by surface plasmon resonance imaging (SPRi) technique. The interaction of gold NPs with mimetic membranes was evaluated by monitoring the variation of reflectivity after several consecutive gold NPs injections on the lipidic membranes prepared on the SPRi biochip. The interaction on the membranes with varied lipidic composition was compared regarding the total surface concentration density of gold NPs adsorbed on them. Then, the interaction of gold and silver NPs with blood proteins was analyzed regarding their kinetic profile of the association/dissociation and dissociation constants (koff). The surface concentration density on the membrane composed of 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine and cholesterol (POPC/cholesterol) was 2.5 times higher than the value found after the injections of gold NPs on POPC only or with dimethyldioctadecylammonium (POPC/DDAB). Regarding the proteins, gold NPs showed preferential binding to fibrinogen resulting in a value of the variation of reflectivity that was 8 times higher than the value found for the other proteins. Differently, silver NPs showed similar interaction on all the tested proteins but with a variation of reflectivity on immunoglobulin G (IgG) 2 times higher than the value found for the other tested proteins.
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  • 文章类型: Journal Article
    表面等离子体共振成像(SPRI)是一种用于可视化折射率变化的强大技术,这使研究人员能够以成像方式观察纳米级物体之间的相互作用。在过去的一段时间里,SPRI的棱镜耦合和非棱镜耦合配置吸引了学者,并发表了许多实验结果。这篇综述描述了SPRI的原理,并详细讨论了棱镜耦合和非棱镜耦合SPRI技术的最新进展,分别。然后,综述了SPRI在生物应用方面的主要进展,包括四个子字段(单元格,病毒,细菌,外泌体,和生物分子)。目的是简要总结SPRI的最新进展,并对SPRI在各个领域的发展进行展望。
    Surface Plasmon Resonance Imaging (SPRI) is a robust technique for visualizing refractive index changes, which enables researchers to observe interactions between nanoscale objects in an imaging manner. In the past period, scholars have been attracted by the Prism-Coupled and Non-prism Coupled configurations of SPRI and have published numerous experimental results. This review describes the principle of SPRI and discusses recent developments in Prism-Coupled and Non-prism Coupled SPRI techniques in detail, respectively. And then, major advances in biological applications of SPRI are reviewed, including four sub-fields (cells, viruses, bacteria, exosomes, and biomolecules). The purpose is to briefly summarize the recent advances of SPRI and provide an outlook on the development of SPRI in various fields.
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