Real-time Investigation of SV40 Large T-antigen Helicase Activity Using Surface Plasmon Resonance

The simian virus 40 (SV40) genome is a model system frequently employed for investigating eukaryotic replication. Large T-antigen (T-ag) is a viral protein responsible for unwinding the SV40 genome and recruiting necessary host factors prior to replication. In addition to duplex unwinding T-ag possesses G-quadruplex DNA helicase activity, the physiological consequence of which is unclear. However, formation of G-quadruplex DNA structures may be involved in genome maintenance and function, and helicase activity to resolve these structures may be necessary for efficient replication. We report the first real-time investigation of SV40 T-ag helicase activity using surface plasmon resonance (SPR). In the presence of ATP, T-ag was observed to bind to immobilized single-stranded DNA, forked duplex DNA, and the human telomeric foldover quadruplex DNA sequence. Inhibition of T-ag duplex helicase activity was observable in real-time and the intramolecular quadruplex was unwound.

Radiative and Non-radiative Surface Plasmon Resonance: Comparison of Real-Time Sensing Performance

Plasmonics - The work presents a surface plasmon resonance sensor able to operate the non-radiative and radiative surface plasmons simultaneously. The optical coupling between photons and surface...     

Real-Time Monitoring of Transferrin-Induced Endocytic Vesicle Formation by Mid-Infrared Surface Plasmon Resonance

We report on the application of surface plasmon resonance (SPR), based on Fourier transform infrared spectroscopy in the mid-infrared wavelength range, for real-time and label-free sensing of transferrin-induced endocytic processes in human melanoma cells. The evanescent field of the mid-infrared surface plasmon penetrates deep into the cell, allowing highly sensitive SPR measurements of dynamic processes occurring at significant cellular depths. We monitored in real-time, infrared reflectivity spectra in the SPR regime from living cells exposed to human transferrin (Tfn). We show that although fluorescence microscopy measures primarily Tfn accumulation in recycling endosomes located deep in the cell's cytoplasm, the SPR technique measures mainly Tfn-mediated formation of early endocytic organelles located in close proximity to the plasma membrane. Our SPR and fluorescence data are very well described by a kinetic model of Tfn endocytosis, suggested previously in similar cell systems. Hence, our SPR data provide further support to the rather controversial ability of Tfn to stimulate its own endocytosis. Our analysis also yields what we believe is novel information on the role of membrane cholesterol in modulating the kinetics of endocytic vesicle biogenesis and consumption.     

Sensitive and Label-Free Detection of DNA by Surface Plasmon Resonance

While an array of technologies based on radioactive labels or luminescent tags are dominant in modern biomedical research on DNA, surface plasmon resonance (SPR) and SPR imaging measurements are sensitive, rapid, and label-free. This review summarizes recent advances in the development of SPR and coupled techniques and their applications in DNA research, including the gene analysis at trace levels and studies of DNA–protein and DNA–drug interactions.     

Nano-gap-Enhanced Surface Plasmon Resonance Sensors

This paper analyzes how dual-mode surface plasmon resonance sensors can be further improved if one were to introduce small (∼20 nm) gaps in the film surface. First, a figure of merit, the sensor’s limit of detection (LOD), is defined in order to optimize the design of the nano-gap sensor. Secondly, the LOD of this design is compared with that of an optimized planar dual-mode design. Through this analysis, it is shown that the LOD of the planar sensor can be improved upon by around a factor of 7 when compared with the nano-gap-enhanced design. Furthermore, with the nano-gap design, the lower wavelength plasmon mode demonstrates remarkably improved selectivity when compared with the conventional sensor. In order to explain these results, the dispersion of each plasmon mode along with the electromagnetic field profiles are modeled and analyzed.     

DNA Detection Based on Localized Surface Plasmon Resonance Spectroscopy of Ag@Au Biocomposite Nanoparticles

Noble metal nanoparticles (NPs) have attracted much attention due to their unique physical and chemical properties such as tunable surface plasmonics, high-efficiency electrochemical sensing, and enhanced fluorescence. We produced two biosensor chips consisting of Ag@Au bimetallic nanoparticles (BNPs) on a carbon thin film by simple RF-sputtering and RF-plasma-enhanced chemical vapor co-deposition. We deposited Au NPs with average size of 4 nm (Au₁ NPs) or 11 nm (Au₂ NPs) on a sensor chip consisting of Ag NPs with mean size of 15 nm, and we investigated the effect of shell size (Au NPs) on the chemical activities of the resulting Ag@Au₁ BNPs and Ag@Au₂ BNPs. We estimated the average size and morphology of Ag@Au BNPs by scanning electron microscopy (SEM) and atomic force microscopy (AFM) images. X-ray diffraction (XRD) patterns revealed that Ag NPs and Au NPs had face-centered cubic (FCC) structure. We studied aging of the biosensor chips consisting of Ag@Au BNPs by localized surface plasmon resonance (LSPR) spectroscopy for up to 3 months. UV–visible aging of the prepared samples indicated that Ag@Au₁ BNPs, which corresponded to Ag NPs covered with smaller Au NPs, were more chemically active than Ag@Au₂ BNPs. Furthermore, we evaluated changes in the LSPR absorption peaks of Ag@Au₁ BNPs and bare Ag NPs in the presence of a DNA primer decamer at fM concentrations, to find that Ag@Au₁ BNPs were more sensitive biosensor chips within a short response time as compared to bare Ag NPs.

     

基于表面等离子体共振和电化学联用的DNA传感器研究进展

表面等离子共振(surface plasmon resonance,SPR)技术旨在检测物体表面附近折射率的变化,其特点是无标记、实时、灵敏和快速,该技术多用于研究分子的相互作用,包括动力学、效率常数和大分子构象变化等。电化学(electrochemical,EC)技术是一项用于定性定量研究电子转移、物质氧化还原、界面吸附等过程的成熟技术,具有简单、低成本和设备小型化的优点。现有的DNA杂交技术,例如光学、电化学或压电转导技术,主要关注于提高DNA杂交检测系统的选择性和灵敏度。传统的SPR在DNA分析方面,由于无法测量折射率的极小变化而在超灵敏检测中的应用受到限制。因此,随着纳米材料的研发和联用技术的飞速发展,SPR与EC联用的生物传感器研究越来越成为人们关注的热点。近年来,关于SPR和EC联用在DNA检测方面的综述鲜有报道。对SPR和EC检测DNA的技术原理、联用方法、应用进展等方面作出了简要的介绍,以期为表面等离子共振和电化学联用的DNA传感器相关研究提供参考。     

Transmission Surface Plasmon Resonance Signal Enhancement via Growth of Gold Nanoparticles on a Gold Grating Surface

We developed a novel technique for increasing the sensitivity of transmission surface plasmon resonance (T-SPR) signals. T-SPR spectroscopy was performed by irradiating, with white light, a gold grating substrate whose surface was nanostructured by growing gold nanoparticles (AuNPs). AuNPs were grown directly on the substrate surface by alcohol reduction and their growth was observed at various stages by UV–visible spectroscopy and standard Kretschmann-type SPR spectroscopy. For comparison, normal gold film with smooth surface was examined under similar condition. The T-SPR results show a possibility of hybrid excitation of both localized and propagating surface plasmon. Significantly, T-SPR spectra of the gold grating substrate obtained during AuNP growth show stronger and narrower peaks in the range 650–800 nm. The maximum T-SPR excitation was at an incident angle of 35°. A layer-by-layer system of 5,10,15,20-tetrakis (1-methyl-4-pyridinio)porphyrin tetra(p-toluenesulfonate) molecules and sodium copper chlorophyllin molecules was used to verify the enhancement of the developed system. We believe that the AuNPs/Au grating for T-SPR devices will provide enhanced signals for detecting nanometer order materials and for high-sensitive sensor applications.     

Visible-Light Photocatalytic Application of Hierarchical Au-ZnO Flower-Rod Heterostructures via Surface Plasmon Resonance

The semiconductor metal oxide, ZnO, is limited to application in photocatalysis because of large bandgap. Metallic nanostructures are utilized to enhance visible-light absorption and improve photocatalytic activity via surface plasmon resonance (SPR). In this work, through a facile thermal decomposition method, the Au nanoparticles (NPs) were successfully loaded on oriented hierarchical ZnO flower-rod (ZFRs) heterostructures, which have a higher areal proportion of exposed active (001) crystal faces. The results indicate that ZFRs are wurtzite phase grown along the [001] direction and the Au NPs spread on the surface of ZFRs in metallic form. It reveals that there is an electronic interaction between Au NPs and ZFRs, and more oxygen molecules are adsorbed on the surface of Au-ZnO flower-rod heterostructures (AZFRs). The AZFRs show a strong absorbance in visible region due to the SPR and enhance the separation of electron–hole pair, resulting in an improvement of photocatalytic activity under visible-light irradiation. During 80 min, the degradation efficiency of rhodamine B for AZFRs-20 is about 3.7 times as the pure ZFRs. Moreover, the AZFRs exhibit predominant photoelectrochemical properties, and the photocurrent can reach 1.2?×?10^?4?A. Finally, the as-prepared photocatalytic device composed of AZFRs based on indium doped tin oxide glass is convenient to recycle without centrifugation.     

Liquid Crystal-Based Surface Plasmon Resonance Biosensor

Plasmonics - Surface plasmon resonance has many applications in designing biosensors. In this paper, an easy fabrication liquid crystal-based surface plasmon resonance (LC-SPR) biosensor is...     

Aluminum-Based Deep-Ultraviolet Surface Plasmon Resonance Sensor

Plasmonics - An aluminum-based deep-ultraviolet surface plasmon resonance (DUV-SPR) sensor is promising for biological applications. Design aspects of a DUV-SPR sensor are here considered by using...     

Ultra-narrow-band Infrared Absorbers Based on Surface Plasmon Resonance

Due to the advantage of improving the sensing performance, narrow-band metamaterial perfect absorbers (MPAs) have attracted much attention in the sensor field. Here, we propose an ultra-narrow-band infrared absorber (UNBIRA) based on localized surface plasmon resonance. The peak absorption of the UNBIRA exceeds 99% with the full width at half maximum (FWHM) of 1.94 nm and 6.32 nm for transverse electric (TE) wave and transverse magnetic (TM) wave in 1.5–1.8 μm. The corresponding Q-factors for TE wave and TM wave are 817 and 266, respectively. When used as an infrared refractive index sensor, the sensitivity of UNBIRA is as high as 1632.5 nm/RIU for TE wave and 1647.5 nm/RIU for TM wave. Accordingly, the figure of merits (FOMs) of 816.2/RIU for TE wave and 260.7/RIU for TM wave are achieved. This UNBIRA possesses a simple geometry structure and an excellent sensing performance, implying a great potential for application of ultra-narrow infrared sensing or detecting.

     

Imaging of Surfaces by Concurrent Surface Plasmon Resonance and Surface Plasmon Resonance-Enhanced Fluorescence

Surface plasmon resonance imaging and surface plasmon induced fluorescent are sensitive tools for surface analysis. However, existing instruments in this area have provided limited capability for concurrent detection, and may be large and expensive. We demonstrate a highly cost-effective system capable of concurrent surface plasmon resonance microscopy (SPRM) and surface plasmon resonance-enhanced fluorescence (SPRF) imaging, allowing for simultaneous monitoring of reflectivity and fluorescence from discrete spatial regions. The instrument allows for high performance imaging and quantitative measurements with surface plasmon resonance, and surface plasmon induced fluorescence, with inexpensive off-the-shelf components.     

Spectral Characteristics of Near-Infrared Surface Plasmon Resonance

Intrinsic properties of surface plasmons (SPs) excited with Kretschmann configuration were analyzed as a function of wavelength, including the propagation length, the penetration depth, the Goos–Hänchen (GH) shift, and the field enhancement. The calculated results indicate that there exists a critical thickness (t _cr) of the gold layer and that the maximum GH shift occurring exactly at the SP resonance wavelength (λ _R) rapidly varies from positive to negative with changing of the gold layer thickness from t?<?t _cr to t?>?t _cr. The maximum field enhancement happens not at λ _R but at a wavelength smaller than λ _R due to the phase retardation between the transmitted and reflected light. Simulations also reveal that a broadband collimated near-infrared beam can simultaneously excite two SPs with different responses to a refractive index (RI) change: the shorter-wavelength SP able to make a small redshift and the longer-wavelength SP capable of yielding a large blueshift. Only the shorter-wavelength SP was experimentally observed and its RI sensitivity was measured to increase from 3,539 nm/RIU at λ _R?=?707.6 nm to 57,143 nm/RIU at λ _R?=?1,398 nm. The SP at λ _R?=?1,013 nm moved to λ _R?=?1,029 nm in response to the saturation adsorption of bovine serum albumin, and the corresponding surface coverage was determined to be Γ?=?1.565 ng/mm² based on a quasilinear dependence of Γ on the resonance wavelength shift (?λ _R) deduced theoretically. Butyrylcholinesterase adsorption from a dilute solution of 10 nM protein in phosphate buffer solution leads to a redshift of ?λ _R?=?10 nm, corresponding to Γ?≈?0.97 ng/mm².     

SPR技术在免疫学研究中的应用

表面等离子共振(Surface Plasmon Resonance,SPR)技术是研究生物分子相互作用的强有力工具之一,该技术使生物分子之间相互作用的实时检测成为可能,并且灵敏度高、无需标记.通过分析传感图谱及分子相互作用的响应值获取分子相互作用的模式和动力学常数等方面的信息,并且获得的信息是能够定性和定量.SPR技术现在已广泛应用于生物、化学、免疫学研究及新药开发等领域.本文主要就SPR技术在免疫学研究中抗体活性检测、抗原表位预测等方面的应用进行了综述.     

表面等离子体激元共振与生物分子相互作用分析

对表面等离子体激元共振（surface plasmon resonance, SPR）的原理和在生物学研究方面的应用进行了综述.这种技术可以直接原位、实时地跟踪生物学实验研究系统,而不需要附加参数如进行标记等手段,具有高敏感性,也可以连续监测吸附或解吸附过程,目前有关的应用涉及到生物学结合分析、动力学及亲和力测定、免疫识别研究、结构与活性研究和核酸研究等多个领域.     

Laccase Activity Assay Using Surface Plasmon Resonance Band of Gold Nanoparticles Formed by Dopamine

Plasmonics - A simple, fast, and sensitive colorimetric technique for determination of laccase activity using dopamine (DA) induced growth of colloidal gold nanoparticles is proposed. It was found...     

Characterization of Glycoprotein Oligosaccharides Using Surface Plasmon Resonance

Surface plasmon resonance was used to characterize the carbohydrate moieties of bovine fetuin. This technique, requiring no sample derivitization or labeling, identified the presence and composition of both N- and O-linked oligosaccharides, using a combination of lectin probes and in situ glycosidase digestion. A complete analysis was achieved using 1.4 μg of pure bovine fetuin, and was fully automated using Pharmacia′s BIAcore. The presence or absence of specific oligosaccharide structures was determined by the binding of a panel of unlabeled lectins. Monosaccharide order and linkages were determined by sequential digestion using a range of specific exoglycosidases. This novel method implementing in situ digestion was achieved using a modification to the BIAcore software, allowing the flow of reagent over the sensor chip to be stopped for variable lengths of time, thereby permitting enzymatic digestion to occur. This technique can be applied to other commercial SPR biosensors currently available. Glycoanalysis by SPR uses approximately 100- to 1000-fold less protein than comparable analyses using alternative techniques such as gel permeation chromatography of released oligosaccharides, labeled lectin binding, or mass spectrometry.     

表面等离子体共振技术在分子生物学中的应用

表面等离子体共振(SPR)技术可以实时、原位地测定生物分子间的相互作用而无需任何标记,可以连续监测吸附和解离过程,并可以进行多组分复合物的相互作用的研究。SPR技术在DNA的复制和转录、DNA的修复、核酸与药物的作用以及肽库和抗体库的筛选等分子生物学领域的应用研究取得了令人瞩目的进展,显示了常规技术无法比拟的优越性。     

Detect the Hybridization of Single-Stranded DNA by Parallel Scan Spectral Surface Plasmon Resonance Imaging

We investigate the ability of a parallel scan spectral surface plasmon resonance imaging system in analyzing hybridization of single-stranded DNA (ssDNA) on microarray. The ssDNA probes are modified by a thiol group and thereby can be immobilized onto the gold film. Hybridization experiments are carried out by using both complementary and noncomplementary sequence to confirm the specificity of interaction. We also demonstrate that the data analysis is very important in reducing the noise and improving the resolution by comparing polynomial fitting method with the combination of polynomial fitting and centroid method. Finally, the results demonstrate that the parallel scan spectral surface plasmon resonance imaging system can be used for high-throughput analysis of the hybridization of the ssDNA.