基于柔性衬底的ZnO葡萄糖酶电极制备及特性

通过水热法在长有ZnO籽晶层的柔性聚酰亚胺(PI)衬底上生长了整齐的ZnO纳米棒,ZnO纳米棒的晶体结构和表面形貌通过X射线衍射(XRD)、扫描电子显微镜(SEM)等进行表征.通过静电吸附方式,将葡萄糖氧化酶(GOx)固定在其表面.分别对GOx及修饰前后的ZnO纳米棒进行了紫外-可见光谱表征,发现修饰后存在ZnO的吸收峰和GOx的特征吸收峰,表明GOx固定在ZnO表面.通过对修饰样品进行傅里叶变换红外(FTIR)光谱测试发现了与GOx相关的吸收峰,这进一步表明GOx仍保持生物活性.最后在循环伏安曲线的测试中,这种在柔性衬底上制备的生物酶电极表现出非常灵敏的电流响应,为制备柔性葡萄糖生物传感器奠定了实验基础.     

Dendrimer-functionalized self-assembled monolayers as a surface plasmon resonance sensor surface

We report here a multistep route for the immobilization of DNA and proteins on chemically modified gold substrates using fourth-generation NH(2)-terminated poly(amidoamine) dendrimers supported by an underlying amino undecanethiol (AUT) self-assembled monolayer (SAM). Bioactive ultrathin organic films were prepared via layer-by-layer self-assembly methods and characterized by fluorescence microscopy, variable angle spectroscopic ellipsometry, atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and attenuated total internal reflection Fourier transform infrared spectroscopy (ATR-FTIR). The thickness of the AUT SAM base layer on the gold substrates was determined to be 1.3 nm from ellipsometry. Fluorescence microscopy and AFM measurements, in combination with analyses of the XPS/ATR-FTIR spectra, confirmed the presence of the dendrimer/biopolymer molecules on the multilayer sensor surfaces. Model proteins, including streptavidin and rabbit immunoglobulin proteins, were covalently attached to the dendrimer layer using linear cross-linking reagents. Through surface plasmon resonance measurements, we found that sensor surfaces containing a dendrimer layer displayed an increased protein immobilization capacity, compared to AUT SAM sensor surfaces without dendrimer molecules. Other SPR studies also revealed that the dendrimer-based surfaces are useful for the sensitive and specific detection of DNA-DNA interactions. Significantly, the multicomponent films displayed a high level of stability during repeated regeneration and hybridization cycles, and the kinetics of the DNA-DNA hybridization process did not appear to be influenced by surface mass transport limiting effects.     

Growth of layered basic zinc acetate in methanolic solutions and its pyrolytic transformation into porous zinc oxide films

Photo-induced reduction of gold and platinum metal salt solutions was carried out using viologen graft copolymerized on low-density polyethylene (LDPE) films and viologen-containing poly(vinylidene fluoride) (PVDF-PVBV) microporous membranes. The effects of the UV irradiation time and concentration of the metal salt solutions on the metal ion reduction process and the resultant metal deposition on the polymeric substrates were investigated. The metal-polymer composites were characterized using X-ray photoelectron spectroscopy (XPS), scanning and transmission electron microscopy (SEM and TEM), atomic force microscopy (AFM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and UV-visible absorption spectroscopy. The amount of metal uptake, the state of the metal, and the size of the metal particles were found to be strongly dependent on the UV irradiation time and the type and concentration of the metal salt solution. The microporous structure and the high viologen content of the PVDF-PVBV membrane constitute an effective matrix for metal ion reduction and preparation of metal nanoparticles.     

Self-assembly, optical behavior, and permeability of a novel capsule based on an azo dye and polyelectrolytes

A novel capsule composed of an azo dye, Congo red (CR), and different polymers, including poly(styrenesulfonate, sodium salt) (PSS), poly(allylamine hydrochloride) (PAH), and poly(diallyldimethylammonium chloride) (PDDA), have been successfully fabricated by the layer-by-layer self-assembly technique. The stepwise linear deposition process was monitored by means of UV-visible absorption measurements. The formation of hollow capsules was verified by confocal laser scanning microscopy (CLSM) and scanning force microscopy (SFM). The resulting hollow PSS/PAH/CR/PDDA capsules displayed a sensitive response to visible light. Optical changes of the hollow capsules prior to and after the photoreaction were investigated in detail by means of UV-visible spectroscopy, CLSM, and SFM. It was found that the photochemical reaction of the assembled hollow capsules depends strongly on the matrix. Qualitative results on the permeability of the hollow capsule walls with CR as one component indicate that the permeability of the walls can be easily photo-controlled at varying irradiation time intervals without addition of external chemicals.     

Atomic force microscopy (AFM)-based nanografting for the study of self-assembled monolayer formation of organophosphonic acids on Al2O3 single-crystal surfaces

Adsorption, stability, and organization kinetics of organophosphonic acids on single-crystalline alumina surfaces were investigated by means of atomic force microscopy (AFM)-based imaging, nanoshaving, and nanografting. AFM friction and phase imaging have shown that chemical etching and subsequent annealing led to heterogeneities on single-crystalline surfaces with (0001) orientation. Self-assembly and stability of octadecylphosphonic acid (ODPA) were shown to be strictly dependent upon the observed heterogeneities of the surface termination, where it was locally shown that ODPA can loosely or strongly bind on different terminations of the crystal surface. Organization kinetics of ODPA was monitored with nanografting on (0001) surfaces. Supported by measurements of surface wettability and diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), it was demonstrated that the lack of organization within the protective adsorbed hexylphosphonic acid (HPA) monolayer on alumina surfaces facilitated the reduced confinement effect during nanografting, such that kinetics information on the organization process of ODPA could be obtained.     

基于层-层自反应的葡萄糖氧化酶有序多层膜电极

以胱胺修饰的金电极为基础电极, 利用席夫碱反应使经高碘酸根氧化的葡萄糖氧化酶在该电极表面进行自身的层-层有序组装. 用电化学交流阻抗法对多层酶膜形成过程的跟踪结果表明, 该多层酶膜的生长是一个逐步形成的均匀过程. 用循环伏安法和I-t曲线法研究了该酶电极对葡萄糖的电催化氧化. 实验结果表明, 当采用羟基二茂铁作为人工电子转移媒介体时, 该酶电极对葡萄糖具有很好的电催化氧化功能. 该传感器制作简便, 响应迅速, 性能稳定, 催化电流与葡萄糖浓度在一定范围内成正比, 并且可以通过控制葡萄糖氧化酶的组装层数来调节该生物传感器的灵敏度与检测限.     

Biocatalytic metal nanopatterning through enzyme-modified microelectrodes

The formation of narrow-size distribution nanomaterials on surfaces in defined patterns is a research area of great interest due to its relevance in many applications such as catalysis, optoelectronics, and sensing devices. Patterning surface with nanostructures has been achieved by numerous techniques including electron-beam lithography, microcontact printing, constructive lithography, and different scanning probe microscopy techniques. Here, we present a different approach by which gold patterns are formed by an enzyme-catalyzed reaction followed by a surface-catalyzed process. Our study takes the advantage of scanning electrochemical microscopy (SECM) where the tip is modified with an enzyme and generates a reductant. The latter participates in an electroless deposition reaction, where AuCl₄⁻ is reduced catalyzed by a Pd surface. The result is local deposition of gold patterns made of nanoparticles as soon as the reductant generated by the tip, i.e., hydroquinone, approaches the Pd surface. Two enzymes were used: glucose oxidase (GOx) and alkaline phosphatase (ALP). The entire process was carefully studied and optimized, which enabled a good control of the patterns formed.

     

Stereoselective Interaction between DNA and Stable Chiral Surfaces Modified with 1,2‐Diphenylethylenediamine Enantiomers

This work described an interesting phenomenon of the stereoselective adsorption behaviors of DNA on stable chiral surfaces which were modified with 1,2‐diphenylethylenediamine enantiomers on gold electrodes. The modification process and electrochemical characterization of the chiral surfaces were measured by cyclic voltammetry (CV). The stereoselective adsorption behaviors of DNA on the two chiral surfaces were investigated via atomic force microscopy (AFM), CV, electrochemical impedance spectroscopy (EIS) and quartz crystal microbalance (QCM). All results confirmed that (1R,2R)‐1,2‐diphenylethylenediamine modified surface had stronger interaction with DNA molecules than (1S,2S)‐1,2‐diphenylethylenediamine modified surface, and the chirality of the surfaces created an different effect on the morphology and adsorption quantity of DNA.     

TAT peptide immobilization on gold surfaces: a comparison study with a thiolated peptide and alkylthiols using AFM, XPS, and FT-IRRAS

A TAT peptide was used to functionalize a gold surface by three different methods: adsorption from solution, microcontact printing, and dip-pen nanolithography (DPN). The composition and structure of the modified gold was characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and Fourier transform -infrared reflection absorption spectroscopy (FT-IRRAS). We used two well-studied alkylthiols, mercaptohexadecanoic acid and 1-octadecanethiol, as a comparison in order to understand the structure of the TAT peptide monolayers prepared by the three methods. AFM studies allowed us to assess the homogeneity after each modification protocol. XPS was used to characterize the chemical composition of the gold surface after each functionalization procedure. The XPS results showed that surfaces modified with the TAT peptide by the three methods exhibit similar surface chemistry. Finally, FT-IRRAS experiments allowed us to conclude that the structure of the alkyl chains of the TAT peptides is fairly disordered and different after each procedure. Regardless of the type of surface functionalization method used, the monolayer of TAT peptide formed on the surface was of "liquidlike" nature.     

Single-molecule chemistry and physics explored by low-temperature scanning probe microscopy

It is well known that scanning probe techniques such as scanning tunnelling microscopy (STM) and atomic force microscopy (AFM) routinely offer atomic scale information on the geometric and the electronic structure of solids. Recent developments in STM and especially in non-contact AFM have allowed imaging and spectroscopy of individual molecules on surfaces with unprecedented spatial resolution, which makes it possible to study chemistry and physics at the single molecule level. In this feature article, we first review the physical concepts underlying image contrast in STM and AFM. We then focus on the key experimental considerations and use selected examples to demonstrate the capabilities of modern day low-temperature scanning probe microscopy in providing chemical insight at the single molecule level.     

Efficient and Facile Fabrication of Glucose Biosensor Based on Electrochemically Etched Porous HOPG Platform

Herein, a uniform porous highly oriented pyrolytic graphite (HOPG) electrode was prepared via diazonium salt assisted electrochemical etching method and firstly utilized to immobilize enzymes for the construction of a high‐performance glucose biosensor. The formation mechanism and morphology structure of the porous HOPG electrode were investigated using atomic force microscopy (AFM), X‐ray photoelectron spectroscopy (XPS) and X‐ray diffraction (XRD) characterizations. The glucose oxidase (GOx) was functionalized with pyrene groups and then immobilized on the porous HOPG substrate through π‐π stacking interactions and hydrogen bonding. As a result, eight times higher oxidation current density can be obtained for a given glucose concentration for the porous HOPG electrode than the pristine one. Detection limit of 5 μM for glucose was achieved for the as‐fabricated biosensor. It was obtained that 78 % biocatalytical activity of GOx can be retained after the pyrene functionalization and 65.7 % one can even be maintained after four weeks, which confirmed the high efficiency and good stability of the as‐prepared biosensor. What's more, it can be anticipated that various other enzymes can be loaded into this porous HOPG platform using the same enzyme modification methodology for the construction of efficient biosensors.     

Characterization of counterion and surface influence on micelle formation using tapping mode atomic force microscopy in air

Cylindrical micelles prepared in aqueous solutions from cationic surfactants octadecyl trimethylammonium (OTA+) or cetyltrimethylammonium (CTA+) and parachlorobenzoate (PCB) counterion were successfully imaged after evaporation of water using tapping mode atomic force microscopy (TM-AFM) onto very smooth gold and glass substrates. With the help of the obtained topography AFM images, it was shown that the micellar structures are preserved on gold substrates after evaporation of the solvent despite the new set of stresses due mainly to capillary forces and dehydration. The influence of the substrate on the resulting micellar morphology observed in air was investigated for these two materials: cylindrical micelles were evidenced as loosely adherent on gold surface in the presence of parachlorobenzoate (PCB) and identical, geometrically speaking, to those known to exist in aqueous solutions. In this situation, topographic AFM images allowed us to determine accurately their geometrical characteristics such as diameter and length in the nanometer range. On the other hand, AFM images obtained in air on glass surfaces revealed micellar structures that are different from those existing in the bulk of the solution. Indeed, bilayer-type micelles with a thickness close to twice the surfactant monomer expected length were observed, indicating that the well-established and strong influence of glass on micelle geometry at the glass/solution interface is maintained after evaporation of water. These results have been analyzed on the basis of positive charge of gold deduced from electrochemical impedance spectroscopy (EIS) and Raman spectroscopy measurements on one hand and of the negative charge of glass on the other hand. Although these results appeal to new theoretical considerations dealing with dynamics of evaporation of micellar solution drops and/or with counterion contributions to macromolecular interactions in aqueous solutions and in air, this new AFM imaging method appears to be the more adequate one to image and measure the micelles formed in the presence of water.     

Amperometric glucose biosensor based on electrodeposition of platinum nanoparticles onto covalently immobilized carbon nanotube electrode

A new amperometric biosensor for glucose was developed based on adsorption of glucose oxidase (GOx) at the gold and platinum nanoparticles-modified carbon nanotube (CNT) electrode. CNTs were covalently immobilized on gold electrode via carbodiimide chemistry by forming amide linkages between carboxylic acid groups on the CNTs and amine residues of cysteamine self-assembled monolayer (SAM). The fabricated GOx/Au_nano/Pt_nano/CNT electrode was covered with a thin layer of Nafion to avoid the loss of GOx in determination and to improve the anti-interferent ability. The immobilization of CNTs on the gold electrode was characterized by quartz crystal microbalance technique. The morphologies of the CNT/gold and Pt_nano/CNT/gold electrodes have been investigated by scanning electron microscopy (SEM), and the electrochemical performance of the gold, CNT/gold, Pt_nano/gold and Pt_nano/CNT/gold electrodes has also been studied by amperometric method. In addition, effects of electrodeposition time of Pt nanoparticles, pH value, applied potential and electroactive interferents on the amperometric response of the sensor were discussed.

The enzyme electrode exhibited excellent electrocatalytic activity and rapid response for glucose in the absence of a mediator. The linear range was from 0.5 to 17.5 mM with correction coefficient of 0.996. The biosensor had good reproducibility and stability for the determination of glucose.     

The dependence of fractal dimension on measuring conditions of scanning probe microscopy

The fractal dimension of sputtered thin film surfaces was determined. Topography was measured using scanning tunneling microscopy (STM) and atomic force microscopy (AFM). It can be shown that measuring conditions have an important influence on the topographic data and the obtained fractal dimension. This influence was investigated systematically. The results of STM- and AFM-measurements have been compared. The results for surfaces imaged with AFM give lower values for the fractal dimension than with STM. Measurements were carried out with a standard cantilever and so the data have been filtered. Dynamic behavior of the measuring system is influenced by scanning speed and loop gain factor. During measurement of topographic signal high scanning speed and low loop gain factor produce a low pass behavior and the fractal dimension will be varied. If there is a disturbance in addition to the topographic signal (e.g. a noise) we found power spectra with more than one linear part. The calculated fractal dimension especially depended on the density of measuring points in the profile. The experimental results were verified by calculated Weierstrass-Mandelbrot functions.     

Development of a Temperature-pulse Enhanced Electrochemical Glucose Biosensor and Characterization of its Stability via Scanning Electrochemical Microscopy

Glucose oxidase (GOx) is an enzyme frequently used in glucose biosensors. As increased temperatures can enhance the performance of electrochemical sensors, we investigated the impact of temperature pulses on GOx that was drop-coated on flattened Pt microwires. The wires were heated by an alternating current. The sensitivity towards glucose and the temperature stability of GOx was investigated by amperometry. An up to 22-fold increase of sensitivity was observed. Spatially resolved enzyme activity changes were investigated via scanning electrochemical microscopy. The application of short (<100 ms) heat pulses was associated with less thermal inactivation of the immobilized GOx than long-term heating.     

Mercury adsorption on gold surfaces employed in the sampling and determination of vaporous mercury: a scanning tunneling microscopy study

To clarify the mechanism of mercury adsorption on gold surfaces thin epitaxial gold films have been exposed to trace amounts of gaseous mercury under laboratory conditions for different periods of time. The changes in the surface morphology of the thin films caused by the exposure have been studied by scanning tunneling microscopy (STM). The evolution of the surface structures with time has been also investigated, in the course of a few days after the exposure. The adsorption of mercury on the gold surfaces has caused drastic changes in the morphology of the surfaces. Pits and islands of 2 to 30 nm in diameter have appeared on the surface, their size and density per unit area depending on the amount of exposure to mercury. The formation of pits and islands followed a certain path of events.     

Mercury adsorption on gold surfaces employed in the sampling and determination of vaporous mercury: a scanning tunneling microscopy study

To clarify the mechanism of mercury adsorption on gold surfaces thin epitaxial gold films have been exposed to trace amounts of gaseous mercury under laboratory conditions for different periods of time. The changes in the surface morphology of the thin films caused by the exposure have been studied by scanning tunneling microscopy (STM). The evolution of the surface structures with time has been also investigated, in the course of a few days after the exposure. The adsorption of mercury on the gold surfaces has caused drastic changes in the morphology of the surfaces. Pits and islands of 2 to 30 nm in diameter have appeared on the surface, their size and density per unit area depending on the amount of exposure to mercury. The formation of pits and islands followed a certain path of events.     

Properties of hybridized DNA arrays on single-crystalline undoped and boron-doped (100) diamonds studied by atomic force microscopy in electrolytes

Properties of hybridized deoxyribonucleic acid (DNA) arrays on single-crystalline undoped and boron-doped diamonds are studied at the microscopic level by atomic force microscopy (AFM) in buffered electrolytic solutions. DNA is linked to diamond via aminodecene molecules (TFAAD) that are attached to undoped diamonds by a photochemical reaction and via nitrophenyl-diazonium molecules attached electrochemically to boron-doped diamonds. Both H-terminated and oxidized diamond surfaces are used in this process. On H-terminated surfaces, AFM measurements detect compact DNA layers. By analyzing phase and height contrast in AFM, a DNA layer height of 76 A is determined on the photochemically functionalized diamonds and a DNA layer height of up to 92 A is determined on the electrochemically functionalized diamonds. Based on the layer thickness, the DNA chains are tilted under the angle of 31 degrees . The morphology of the DNA layers exhibits long-range (30-50 nm) undulations of 20 A in height and a nanoroughness of 8 A. Using Hertz's model for calculating the contact area of the AFM tip on a DNA layer and a geometrical model of DNA arrangement on diamond yields the DNA density on diamonds of 6 x 10(12) cm(-2) on both photochemically and electrochemically functionalized diamonds. The structure of these dense DNA layers is not significantly influenced by variations in buffer salinity of 1-300 mM NaCl. DNA molecules can be removed from the diamond surface by contact-mode AFM with forces >or= 45 nN and >or= 76 nN on photochemically and electrochemically functionalized diamonds, respectively, indicating that DNA is bonded covalently and stronger on diamond than on gold substrates. The DNA arrangement and bonding strength are similar on oxidized diamond surfaces when using an electrochemical process. On oxidized surfaces after photochemical processing, DNA is bonded noncovalently as deduced from the removal force < 6 nN. The presence of hybridized DNA as well as the selective removal of DNA by AFM scanning are corroborated by fluorescence microscopy.     

Surface grafted sulfobetaine polymers via atom transfer radical polymerization as superlow fouling coatings

One of the sulfobetaine methacrylate (SBMA) monomers, N-(3-sulfopropyl)-N-(methacryloxyethyl)-N,N-dimethylammonium betaine, was polymerized onto initiator-covered gold surfaces using atom transfer radical polymerization (ATRP) to form uniform polymer brushes. Self-assembled monolayers (SAMs) with ATRP initiators were characterized by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The thickness of grafted poly(SBMA) films was measured by ellipsometry. Fibrinogen adsorption on poly(SBMA) grafted surfaces was measured with a surface plasmon resonance (SPR) sensor. Two approaches were compared to graft ATRP initiators onto gold surfaces for surface polymerization and subsequent protein adsorption on these polymer grafted surfaces. The first was to prepare a SAM from omega-mercaptoundecyl bromoisobutyrate onto a gold surface. Superlow fouling surfaces with well-controlled poly(SBMA) brushes were achieved using this approach (e.g., fibrinogen adsorption <0.3 ng/cm2). The second approach was to react bromoisobutyryl bromide with a hydroxyl-terminated SAM on a gold surface. Although protein adsorption decreased as the density of surface initiators increased, the surface prepared using the second approach was not able to achieve as low protein adsorption as the first approach. Key parameters to achieve superlow fouling surfaces were studied and discussed.     

Characterization of single- and double-stranded DNA on gold surfaces

Single- and double-stranded deoxy ribonucleic acid (DNA) molecules attached to self-assembled monolayers (SAMs) on gold surfaces were characterized by a number of optical and electronic spectroscopic techniques. The DNA-modified gold surfaces were prepared through the self-assembly of 6-mercapto-1-hexanol and 5'-C(6)H(12)SH -modified single-stranded DNA (ssDNA). Upon hybridization of the surface-bound probe ssDNA with its complimentary target, formation of double-stranded DNA (dsDNA) on the gold surface is observed and in a competing process, probe ssDNA is desorbed from the gold surface. The competition between hybridization of ssDNA with its complimentary target and ssDNA probe desorption from the gold surface has been investigated in this paper using X-ray photoelectron spectroscopy, chronocoulometry, fluorescence, and polarization modulation-infrared reflection absorption spectroscopy (PM-IRRAS). The formation of dsDNA on the surface was identified by PM-IRRAS by a dsDNA IR signature at approximately 1678 cm(-)(1) that was confirmed by density functional theory calculations of the nucleotides and the nucleotides' base pairs. The presence of dsDNA through the specific DNA hybridization was additionally confirmed by atomic force microscopy through colloidal gold nanoparticle labeling of the target ssDNA. Using these methods, strand loss was observed even for DNA hybridization performed at 25 degrees C for the DNA monolayers studied here consisting of attachment to the gold surfaces by single Au-S bonds. This finding has significant consequence for the application of SAM technology in the detection of oligonucleotide hybridization on gold surfaces.