扫描探针显微术在巯醇自组装单分子膜纳米刻蚀中的应用

介绍了近十年来扫描探针显微术(SPM)在巯醇自组装单分子膜纳米刻蚀中的应用. 依据扫描探针的工作原理, 依次讨论了扫描隧道显微镜、原子力显微镜和导电原子力显微镜的工作特点和适用范围. 同时也讨论了自组装单分子膜纳米刻蚀术在生物分子传感器、超高密度信息存储等领域的应用前景.     

扫描探针刻蚀技术可控构建牛血清白蛋白纳米结构

利用Dip-pen纳米刻蚀技术(简称DPN技术)在云母基底上构建出形状、尺寸可控的牛血清白蛋白(BSA)纳米结构.考察了针尖接触基底时间及针尖下行距离对构建的牛血清白蛋白纳米结构的影响.较长的针尖-基底接触时间及较深的下行距离可以沉积更多的牛血清白蛋白分子,构建牛血清白蛋白纳米结构的形状除了与墨水分子的本身性能有关,还与墨水-基底的相互作用有关.这些形状及尺寸可控的蛋白质纳米结构可以作为模板,进行金属、半导体等其它材料的组装,有望用于制造光电纳米器件及生物纳米器件.     

Polymer Nanostructures Made by Scanning Probe Lithography: Recent Progress in Material Applications

Scanning probe lithography (SPL) is a series of techniques that utilizes a scanning probe or an array of probes for surface patterning. Recent developments of new material systems and patterning approaches have made SPL a promising, low‐cost, bench‐top, and versatile tool for fabricating various polymer nanostructures, with extraordinary importance in physical sciences, life sciences and nanotechnology. This feature article highlights the recent progress in four material applications: polymer resists, polymeric carriers for patterning functional materials, electronically active polymers and polymer brushes for tailoring surface morphology and functionality. An overview of future possibilities, with regard to challenges and opportunities in this field, is given at the end of the paper.     

Deformation of elastomeric pyramid pen arrays in cantilever‐free scanning probe lithography

Cantilever‐free scanning probe lithography (SPL) employs soft elastomeric pen arrays to deliver material or energy to a surface to achieve a high‐resolution, high‐throughput, and low‐cost nanopatterning. In particular, microscale elastomeric pyramid pen arrays are often adopted as the cantilever‐free architecture owing to their distinct structural and mechanical properties. To better understand the mechanical behavior of the elastomeric pyramid pen array during the lithographic printing process, we numerically investigate the compression of an elastomeric pyramid array in a nonadhesive and frictionless contact with a rigid substrate. Simple scaling laws of the width of the contact surface with respect to the compression displacement and force are found and compared with previous models and experiments. By changing the interpyramid distance or the thickness of the base of the pyramid array, increasing deviations from the established scaling laws are observed and explained. Furthermore, we demonstrate that the unique morphology of a compressed pyramid primarily determines the unusual shape of the features fabricated by a specific cantilever‐free SPL technique. © 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2018 , 56, 731–738     

Fabrication of thermosensitive polymer nanopatterns through chemical lithography and atom transfer radical polymerization

Micro- and nanopatterns of thermosensitive poly(N-isopropylacrylamide) brush on gold substrate were prepared by using chemical lithography combined with surface-initiated atom transfer radical polymerization. Self-assembled monolayers of 4'-nitro-1, 1'-biphenyl-4-thiol were structured by chemical lithography which produced cross-linked 4'-amino-1,1'-biphenyl-4-thiol monolayer within a nitro-terminated matrix. The terminal amino groups in monolayers were bounded with the surface initiator bromoisobutyryl bromide. After polymerization, the smallest size can reach to 70-nm line width and dots. The thermosensitivity of poly(N-isopropylacrylamide) brushes is demonstrated by contact angle measurement and fluid atomic force microscopy. This fabrication approach allows creating spatially defined polymer patterns and provides a simple and versatile method to construct complex micro- and nanopatterned polymer brushes with spatial and topographic control in a single step.     

Patterning biomolecules with a water-soluble release and protection interlayer

We report on a general lithography method for high-resolution biomolecule patterning with a bilayer resist system. Biomolecules are first immobilized on the surface of a substrate and covered by a release-and-protection interlayer of water-soluble polymer. Patterns can then be obtained by lithography with a spin-coated resist layer in a conventional way and transferred onto the substrate by reactive ion etching. Afterward, the resist layer is removed by dissolution in water. To demonstrate a high-resolution patterning, soft UV nanoimprint lithography has been used to produce high-density dot arrays of poly-(L-lysine) molecules on a glass substrate. Both fluorescence images and cell proliferation behaviors on such a patterned substrate have shown evidence of improved stability of biomolecule immobilization comparing to that obtained by microcontact printing techniques.     

Hybrid protein-lipid patterns from aluminum templates

An aqueous aluminum liftoff process suitable for fabrication of hybrid patterns of protein and supported lipid membrane on silica surfaces is described. Patterned aluminum thin films, which can be produced by conventional optical or electron beam lithography, are employed as sacrificial protecting layers to define the geometry of the protein-lipid patterns. The aluminum is lifted off in a mildly basic aqueous solution, which preserves the integrity of bound protein layers. The newly exposed substrate can then be filled with supported membrane by exposure to an aqueous vesicle suspension. The final substrate consists of patterned protein and lipid membranes with spatial resolution determined by aluminum patterns, down to 200 nm line widths in this case. Inorganic surfaces were characterized by atomic force microscopy and X-ray photoelectron spectroscopy while supported bilayers and protein patterns were characterized by epifluorescence microscopy.     

原子力显微镜研究高分子超薄膜结晶机理及功能化调控

近十年来,随着功能高分子单晶(含单层或寡层片晶)工程及应用研究的不断深入,除了纳米尺度结晶形貌的表征以外,多功能原子力显微镜还被用于研究分子结构、结晶条件和后处理条件对功能高分子晶体性能(电、热、光、磁等)的影响,进一步还可采用扫描探针加工技术(机械刻蚀、电致刻蚀和热致刻蚀等)对其性能进行调控以构筑功能化聚集态结构和微图案.另一方面,超薄膜中单层或寡层片晶可为研究高分子结晶提供合适的模型体系,与原子力显微镜相结合,不但可以原位、实空间、高分辨地研究高分子的成核与生长过程(生长形态演变和生长动力学),还可以用于研究亚稳态折叠链片晶厚度和形态随热处理温度与时间的演化,从而加深对片晶内有序差异、片晶增厚与熔融行为和自诱导成核的认识.     

Patterned Growth of Vertically Aligned Polypyrrole Nanowire Arrays

A facile strategy for preparing vertically aligned polypyrrole (PPy) nanoarrays with precisely controlled density and quantity is presented. The method involves two steps: (1) the fabrication of the patterned substrate via electron beam lithography and (2) the controlled growth of PPy nanowires via electrochemical polymerization on the patterned substrate. The electrical property of a single PPy nanowire is investigated via in situ conducting probe atomic force microscopy.     

Electric-field-controlled flow in nanoscale-thin wetting films

A novel nanofluidic system based on electroosmotic flow in nanoscale-thin aqueous wetting films is reported. The water films formed spontaneously on mica substrates in a saturation humidity environment. The film thickness was determined to be a few tens of nanometers by optical interference and fluorescence intensity measurements and was consistent with a theoretical evaluation of the thickness of a film based on the competing forces of electrostatic repulsion and capillary pressure. Lateral flow was induced by applying a dc electric field tangential to the film and characterized by tracking the position of a fluorescent probe. The mobilities of the thin fluid layer and the flow marker were lower than the predictions of the electrokinetic theory, which may be a result of adsorption of the fluorescent molecules to the mica. Confinement of the film to two-dimensional "channels" was achieved by microcontact printing of patterned hydrophobic monolayers onto the substrate. This system has the advantage of simple and inexpensive fabrication in comparison to nanofluidic devices made by traditional lithography techniques.     

Nanostructures of functionalized gold nanoparticles prepared by particle lithography with organosilanes

Periodic arrays of organosilane nanostructures were prepared with particle lithography to define sites for selective adsorption of functionalized gold nanoparticles. Essentially, the approach for nanoparticle lithography consists of procedures with two masks. First, latex mesospheres were used as a surface mask for deposition of an organosilane vapor, to produce an array of holes within a covalently bonded, organic thin film. The latex particles were readily removed with solvent rinses to expose discrete patterns of nanosized holes of uncovered substrate. The nanostructured film of organosilanes was then used as a surface mask for a second patterning step, with immersion in a solution of functionalized nanoparticles. Patterned substrates were fully submerged in a solution of surface-active gold nanoparticles coated with 3-mercaptopropyltrimethoxysilane. Regularly shaped, nanoscopic areas of bare substrate produced by removal of the latex mask provided sites to bind silanol-terminated gold nanoparticles, and the methyl-terminated areas of the organosilane film served as an effective resist, preventing nonspecific adsorption on masked areas. Characterizations with atomic force microscopy demonstrate the steps for lithography with organosilanes and functionalized nanoparticles. Patterning was accomplished for both silicon and glass substrates, to generate nanostructures with periodicities of 200-300 nm that match the diameters of the latex mesospheres of the surface masks. Nanoparticles were shown to bind selectively to uncovered, exposed areas of the substrate and did not attach to the methyl-terminal groups of the organosilane mask. Billions of well-defined nanostructures of nanoparticles can be generated using this high-throughput approach of particle lithography, with exquisite control of surface density and periodicity at the nanoscale.     

基于粒子辅助水滴模板法制备仿生复眼结构的研究

将微粒“皮克林乳化效应”(Pickering emulsions)和水滴模板法(breath figure method)有机结合,探索通过建立粒子辅助的水滴模板法,实现纳米粒子在蜂窝状多孔膜内壁的自组装复合,构建微纳复合的多级仿生结构.并进一步利用聚二甲基硅氧烷(PDMS)复制转移技术,获得类似于复眼结构的多级微纳复合界面仿生结构.     

Mathematical modeling of the action of biosensor possessing variable parameters

Electrochemical biosensor containing flat semi-permeable membrane covering enzyme-containing layer has been investigated. Mathematical modeling of the action modes of electrochemical biosensors with outer diffusion membrane was performed. Operation of the biosensor under the conditions when the permeability of the membrane and the activity of the biocatalytic layer depend on the parameters of the probe has been examined. The pH and temperature were selected as the main parameters which often affect the action of biosensors. A set of parameters was selected when the biosensor operates in kinetic and diffusion modes of action. The response time of the biosensor was shown to be sensitive to the mode of the biosensor action especially in the boundary region of the biosensor action. The linearity of the biosensor (the linear dependence of the biosensor response on the substrate concentration) in the deep diffusion mode can be increased by several magnitudes, whereas the response time increases only several times.     

Mechanosensing using drag force for imaging soft biological membranes

We investigate physical processes taking place during nanoscale mechanosensing of soft biological membranes in liquid environments. Examples include tapping mode imaging by atomic force microscope (AFM) and microscopy based on the Brownian motion of a nanoparticle in an optical-tweezers-controlled trap. The softness and fluidity of the cellular membrane make it difficult to accurately detect (i.e., image) the shape of the cell using traditional mechanosensing methods. The aim of the reported work is to theoretically evaluate whether the drag force acting on the nanoscale mechanical probe due to a combined effect of intra- and extracellular environments can be exploited to develop a new imaging mode suitable for soft cellular interfaces. We approach this problem by rigorous modeling of the fluid mechanics of a complex viscoelastic biosystem in which the probe sensing process is intimately coupled to the membrane biomechanics. The effects of the probe dimensions and elastic properties of the membrane as well as intra- and extracellular viscosities are investigated in detail to establish the structure and evolution of the fluid field as well as the dynamics of membrane deformation. The results of numerical simulations, supported by predictions of the scaling analysis of forces acting on the probe, suggest that viscous drag is the dominant force dictating the probe dynamics as it approaches a biological interface. The increase in the drag force is shown to be measurable, to scale linearly with an increase in the viscosity ratio of the fluids on either side of the membrane, and to be inversely proportional to the probe-to-membrane distance. This leads to the postulation of a new strategy for lipid membrane imaging by AFM or other mechanosensing methods using a variation in the maximum drag force as an indicator of the membrane position.     

Genotyping by alkaline dehybridization using graphically encoded particles

This work describes a nonenzymatic, isothermal genotyping method based on the kinetic differences exhibited in the dehybridization of perfectly matched (PM) and single-base mismatched (MM) DNA duplexes in an alkaline solution. Multifunctional encoded hydrogel particles incorporating allele-specific oligonucleotide (ASO) probes in two distinct regions were fabricated by using microfluidic-based stop-flow lithography. Each particle contained two distinct ASO probe sequences differing at a single base position, and thus each particle was capable of simultaneously probing two distinct target alleles. Fluorescently labeled target alleles were annealed to both probe regions of a particle, and the rate of duplex dehybridization was monitored by using fluorescence microscopy. Duplex dehybridization was achieved through an alkaline stimulus using either a pH step function or a temporal pH gradient. When a single target probe sequence was used, the rate of mismatch duplex dehybridization could be discriminated from the rate of perfect match duplex dehybridization. In a more demanding application in which two distinct probe sequences were used, we found that the rate profiles provided a means to discriminate probe dehybridizations from both of the two mismatched duplexes as well as to distinguish at high certainty the dehybridization of the two perfectly matched duplexes. These results demonstrate an ability of alkaline dehybridization to correctly discriminate the rank hierarchy of thermodynamic stability among four sets of perfect match and single-base mismatch duplexes. We further demonstrate that these rate profiles are strongly temperature dependent and illustrate how the sensitivity can be compensated beneficially by the use of an actuating gradient pH field.     

Imaging Local Proton Fluxes through a Polycarbonate Membrane by Using Scanning Electrochemical Microscopy and Functionalized Alkanethiols

A new application of scanning electrochemical microscopy (SECM) to probe the transport of protons through membranes is described. Herein, a probe ultramicroelectrode (UME) is modified with a self‐assembled monolayer (SAM) of 11‐mercaptoundecanoic acid to qualitatively image areas within different pH regions above a track‐etched membrane. The current response of the modified electrode in the presence of potassium hexacyanoferrate as electroactive component is different in acidic and alkaline solutions. Depending on the pH value of the solution, the SAM‐covered electrode exposes either a neutral or a negatively charged insulating monolayer at pH 3 or 7, respectively, which leads to an increase/decrease in the faradaic current due to electrostatic interactions between the neutral/charged surface and the charged redox mediator. Therefore, local pH changes in the close vicinity of a membrane‐like substrate lead to different current responses recorded at the tip electrode when scanning above the surface.     

Nanoreactors for studying single nanoparticle coarsening

The ability to observe intermediate structures as part of coarsening processes that lead to the formation of single nanoparticles (NPs) is important in gaining fundamental insight pertaining to nanostructure growth. Here, we use scanning probe block copolymer lithography (SPBCL) to create "nanoreactors" having attoliter volumes, which confine Au NP nucleation and growth to features having diameters <150 nm on a substrate. With this technique, one can use in situ TEM to directly observe and study NP coarsening and differentiate Ostwald ripening from coalescence processes. Importantly, the number of metal atoms that can engage in coarsening can be controlled with this technique, and TEM "snapshots" of particle growth can be taken. The size of the resulting nanostructures can be controlled in the 2-10 nm regime.     

A wireless pH sensor using magnetoelasticity for measurement of body fluid acidity.

The determination of body fluid acidity using a wireless magnetoelastic pH-sensitive sensor is described. The sensor was fabricated by casting a layer of pH-sensitive polymer on a magnetoelastic ribbon. In response to an externally applied time-varying magnetic field, the magnetoelastic sensor mechanically vibrates at a characteristic frequency that is inversely dependent upon the mass of the pH polymer film, which varies as the film swells and shrinks in response to pH. As the magnetoelastic sensor is magnetostrictive, the mechanical vibrations of the sensor launch magnetic flux that can be detected remotely using a pickup coil. The sensor can be used for direct measurements of body fluid acidity without a pretreatment of the sample by using a filtration membrane. A reversible and linear response was obtained between pH 5.0 and 8.0 with a measurement resolution of pH 0.1 and a slope of 0.2 kHz pH(-1). Since there are no physical connections between the sensor and the instrument, the sensor can be applied to in vivo and in situ monitoring of the physiological pH and its fluctuations.     

A Micro Planar Ampehometric Glucose Sensor Using an Isfet as a Reference Electrode

Abstract

A very small glucose sensor has been realized, which consists of a gold working electrode with a glucose oxidase immobilized membrane on it, and a gold counter electrode, all made on a sapphire substrate. By using the pH sensitive ISFET as a reference electrode, the potential for a solution, whose pH is constant, can be measured and irreversible metal electrodes, such as gold or platinum, can be used as working electrode and counter electrode. The sensor is very suitable for miniaturizing and mass production, because the Integrated Circuit (IC) fabrication process can be applied. The glucose oxidase immobilized membrane was also deposited by a lift off method, one of the IC processes. A glucose concentration, from 1 to 100 mg/dl, was measured with good linear current output.     

Development of a BODIPY-based ratiometric fluorescent probe for hypochlorous acid and its application in living cells

A BODIPY-based ratiometric fluorescent probe for HOCl has been designed based on the transduction of thioether to sulfoxide function. This probe features a marked absorption and emission blue-shift upon the HOCl-promoted rapid transduction, enabling the highly selective and ratiometric detection. In addition, the probe works excellently within a wide pH range of 4–10, addressing the existing pH dependency issue. Living cells studies demonstrate that the probe is cell membrane permeable and can be employed successfully to image endogenous HOCl generation in macrophage cells.