Fabrication of nanopores in a 100-nm thick Si3N4 membrane

Textured alumina films have been used to fabricate nanoscale pores in Si3N4 membranes. A few nanometer-thick alumina layer was used as a masking material for nanopore fabrication, and the pattern was transferred into a 100-nm thick, 200 microm x 200 microm Si3N4 membrane by reactive ion etching (RIE). The nanopores were found to be concentrated in a approximately 150-microm diameter region at the center of the membrane.     

Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores

Most experiments on nanopores have concentrated on the pore-forming protein α-haemolysin (αHL) and on artificial pores in solid-state membranes. While biological pores offer an atomically precise structure and the potential for genetic engineering, solid-state nanopores offer durability, size and shape control, and are also better suited for integration into wafer-scale devices. However, each system has significant limitations: αHL is difficult to integrate because it relies on delicate lipid bilayers for mechanical support, and the fabrication of solid-state nanopores with precise dimensions remains challenging. Here we show that these limitations may be overcome by inserting a single αHL pore into a solid-state nanopore. A double-stranded DNA attached to the protein pore is threaded into a solid-state nanopore by electrophoretic translocation. Protein insertion is observed in 30-40% of our attempts, and translocation of single-stranded DNA demonstrates that the hybrid nanopore remains functional. The hybrid structure offers a platform to create wafer-scale device arrays for genomic analysis, including sequencing.     

Periodic arrays of deep nanopores made in silicon with reactive ion etching and deep UV lithography

We report on the fabrication of periodic arrays of deep nanopores with high aspect ratios in crystalline silicon. The radii and pitches of the pores were defined in a chromium mask by means of deep UV scan and step technology. The pores were etched with a reactive ion etching process with SF(6), optimized for the formation of deep nanopores. We have realized structures with pitches between 440 and 750?nm, pore diameters between 310 and 515?nm, and depth to diameter aspect ratios up to 16. To the best of our knowledge, this is the highest aspect ratio ever reported for arrays of nanopores in silicon made with a reactive ion etching process. Our experimental results show that the etching rate of the nanopores is aspect-ratio-dependent, and is mostly influenced by the angular distribution of the etching ions. Furthermore we show both experimentally and theoretically that, for sub-micrometer structures, reducing the sidewall erosion is the best way to maximize the aspect ratio of the pores. Our structures have potential applications in chemical sensors, in the control of liquid wetting of surfaces, and as capacitors in high-frequency electronics. We demonstrate by means of optical reflectivity that our high-quality structures are very well suited as photonic crystals. Since the process studied is compatible with existing CMOS semiconductor fabrication, it allows for the incorporation of the etched arrays in silicon chips.     

Nanoparticle transport in conical-shaped nanopores

This report presents a fundamental study of nanoparticle transport phenomena in conical-shaped pores contained within glass membranes. The electrophoretic translocation of charged polystyrene (PS) nanoparticles (80- and 160-nm-radius) was investigated using the Coulter counter principle (or "resistive-pulse" method) in which the time-dependent nanopore current is recorded as the nanoparticle is driven across the membrane. Particle translocation through the conical-shaped nanopore results in a direction-dependent and asymmetric triangular-shaped resistive pulse. Because the sensing zone of conical-shaped nanopores is localized at the orifice, the translocation of nanoparticles through this zone is very rapid, resulting in pulse widths of ~200 μs for the nanopores used in this study. A linear dependence between translocation rate and nanoparticle concentration was observed from 10(7) to 10(11) particles/mL for both 80- and 160-nm-radius particles, and the magnitude of the resistive pulse scaled approximately in proportion to the particle volume. A finite-element simulation based on continuum theory to compute ion fluxes was combined with a dynamic electric force-based nanoparticle trajectory calculation to compute the position- and time-dependent nanoparticle velocity as the nanoparticle translocates through the conical-shaped nanopore. The computational results were used to compute the resistive pulse current-time response for conical-shaped pores, allowing comparison between experimental and simulated pulse heights and translocation times. The simulation and experimental results indicate that nanoparticle size can be differentiated based on pulse height, and to a lesser extent based on translocation time.     

Aligned nanowire growth using lithography-assisted bonding of a polycarbonate template for neural probe electrodes

This research presents a fabrication method of vertically aligned nanowires on substrates using lithography-assisted template bonding (LATB) towards developing highly efficient electrodes for biomedical applications at low cost. A polycarbonate template containing cylindrical nanopores is attached to a substrate and the nanopores are selectively opened with a modified lithography process. Vertically aligned nanowires are grown by electrochemical deposition through these open pores on polyimide film and silicon substrates. The process of opening the nanopores is optimized to yield uniform growth of nanowires. The morphological, crystalline, and electrochemical properties of the resulting vertically aligned nanowires are discussed using scanning electron microscopy (SEM), x-ray diffraction (XRD), and electrochemical analysis tools. The potential application of this simple and inexpensive fabrication technology is discussed in the development of neural probe electrodes.     

Fabrication of nanofluidic biochips with nanochannels for applications in DNA analysis

With the development of nanotechnology, great progress has been made in the fabrication of nanochannels. Nanofluidic biochips based on nanochannel structures allow biomolecule transport, bioseparation, and biodetection. The domain applications of nanofluidic biochips with nanochannels are DNA stretching and separation. In this Review, the general fabrication methods for nanochannel structures and their applications in DNA analysis are discussed. These representative fabrication approaches include conventional photolithography, interference lithography, electron-beam lithography, nanoimprint lithography and polymer nanochannels. Other nanofabrication methods used to fabricate unique nanochannels, including sub-10-nm nanochannels, single nanochannels, and vertical nanochannels, are also mentioned. These nanofabrication methods provide an effective way to form nanoscale channel structures for nanofluidics and biosensor devices for DNA separation, detection, and sensing. The broad applications of nanochannels and future perspectives are also discussed.     

Voltage-gated ion transport through semiconducting conical nanopores formed by metal nanoparticle-assisted plasma etching

Nanopores with conical geometries have been found to rectify ionic current in electrolytes. While nanopores in semiconducting membranes are known to modulate ionic transport through gated modification of pore surface charge, the fabrication of conical nanopores in silicon (Si) has proven challenging. Here, we report the discovery that gold (Au) nanoparticle (NP)-assisted plasma etching results in the formation of conical etch profiles in Si. These conical profiles result due to enhanced Si etch rates in the vicinity of the Au NPs. We show that this process provides a convenient and versatile means to fabricate conical nanopores in Si membranes and crystals with variable pore-diameters and cone-angles. We investigated ionic transport through these pores and observed that rectification ratios could be enhanced by a factor of over 100 by voltage gating alone, and that these pores could function as ionic switches with high on-off ratios of approximately 260. Further, we demonstrate voltage gated control over protein transport, which is of importance in lab-on-a-chip devices and biomolecular separations.     

Nanochannel system fabricated by MEMS microfabrication and atomic force microscopy

A silicon nanochannel system with integrated transverse electrodes was designed and fabricated by combining micro-electro-mechanical systems (MEMS) micromachining and atomic force microscopy (AFM)-based nanolithography. The fabrication process began with the patterning of microscale reservoirs and electrodes on an oxidised silicon chip using conventional MEMS techniques. A nanochannel, approximately 30?[micro sign]m long with a small semi-circular cross-sectional area of 20?nm × 200?nm, was then mechanically machined on the oxide surface between the micro reservoirs by applying AFM nanolithography with an all-diamond probe. Anodic bonding was used to seal off the nanochannel with a matching Pyrex cover. Continuous flow in the nanochannel was verified by pressurising a solution of fluorescein isothiocyanate in ethanol through the nanochannel in a vacuum chamber. It was further demonstrated by translocating negatively charged nanobeads (diameter approximately 20?nm) through the nanochannel by using an external DC electric field. The passage of the nanobeads caused a sharp increase in the transverse electrical conductivity of the nanochannel.     

Controlling molecular transport through nanopores

Nanopores are emerging as powerful tools for the detection and identification of macromolecules in aqueous solution. In this review, we discuss the recent development of active and passive controls over molecular transport through nanopores with emphasis on biosensing applications. We give an overview of the solutions developed to enhance the sensitivity and specificity of the resistive-pulse technique based on biological and solid-state nanopores.     

Characterization of nanostructured surfaces generated by reconstitution of the porin MspA from Mycobacterium smegmatis

Nanostructures with long-term stability at the surface of gold electrodes are generated by reconstituting the porin MspA from Mycobacterium smegmatis into a specially designed monolayer of long-chain lipid surfactant on gold. Tailored surface coverage of gold electrodes with long-chain surfactants is achieved by electrochemically assisted deposition of organic thiosulfates (Bunte salts). The subsequent reconstitution of the octameric-pore MspA is guided by its extraordinary self-assembling properties. Importantly, electrochemical reduction of copper(II) yields copper nanoparticles within the MspA nanopores. Electrochemical impedance spectroscopy, reflection electron microscopy, and atomic force microscopy (AFM) show that: 1) the MspA pores within the self-assembled monolayer (SAM) are monodisperse and electrochemically active, 2) MspA reconstitutes in SAMs and with a 10-nm thickness, 3) AFM is a suitable method to detect pores within SAMs, and 4) the electrochemical reduction of Cu2+ to Cu0 under overpotential conditions starts within the MspA pores.     

Morphological evolution of porous nanostructures grown from a single isolated anodic alumina nanochannel

Porous anodic aluminum oxide (AAO) membranes have been widely used as templates for growing nanomaterials because of their ordered nanochannel arrays with high aspect ratio and uniform pore diameter. However, the intrinsic growth behavior of an individual AAO nanochannel has never been carefully studied for the lack of a means to fabricate a single isolated anodic alumina nanochannel (SIAAN). In this study, we develop a lithographic method for fabricating a SIAAN, which grows into a porous hemispherical structure with its pores exhibiting fascinating morphological evolution during anodization. We also discover that the mechanical stress affects the growth rate and pore morphology of AAO porous structures. This study helps reveal the growth mechanism of arrayed AAO nanochannels grown on a flat aluminum surface and provides insights to help pave the way to altering the geometry of nanochannels on AAO templates for the fabrication of advanced nanocomposite materials.     

Study of nanopores ordering in anodic aluminum oxide templates achieved by three step process

Enhancement of naturally-occurred self ordering nanopores in anodic aluminum oxide membrane by performing three-step anodic oxidation process has been reported. Naturally-occurred self ordering of nanopores in anodic aluminum oxide membrane has brought it into the applications of template for fabrication of nanoscale materials. Three-step anodic oxidation method was used to achieve self-ordering of nanopores. Effect of duration of first and second steps on the ordering of nanopores was investigated. The current-time curves recorded during anodization elucidate an almost same behavior for all three steps. Scanning electron micrographs show hexagonally arranged 45 nm pores in a manner which contribute into the formation of highly ordered areas, called domains. Larger ones are clearly observed over the surface, for samples with longer first and second anodization steps.     

Fabrication of solid-state nanopores with single-nanometre precision

Single nanometre-sized pores (nanopores) embedded in an insulating membrane are an exciting new class of nanosensors for rapid electrical detection and characterization of biomolecules. Notable examples include alpha-hemolysin protein nanopores in lipid membranes and solid-state nanopores in Si3N4. Here we report a new technique for fabricating silicon oxide nanopores with single-nanometre precision and direct visual feedback, using state-of-the-art silicon technology and transmission electron microscopy. First, a pore of 20 nm is opened in a silicon membrane by using electron-beam lithography and anisotropic etching. After thermal oxidation, the pore can be reduced to a single-nanometre when it is exposed to a high-energy electron beam. This fluidizes the silicon oxide leading to a shrinking of the small hole due to surface tension. When the electron beam is switched off, the material quenches and retains its shape. This technique dramatically increases the level of control in the fabrication of a wide range of nanodevices.     

Rapid and precise scanning helium ion microscope milling of solid-state nanopores for biomolecule detection

We report the formation of solid-state nanopores using a scanning helium ion microscope. The fabrication process offers the advantage of high sample throughput along with fine control over nanopore dimensions, producing single pores with diameters below 4 nm. Electronic noise associated with ion transport through the resultant pores is found to be comparable with levels measured on devices made with the established technique of transmission electron microscope milling. We demonstrate the utility of our nanopores for biomolecular analysis by measuring the passage of double-strand DNA.     

Ion-beam pore opening of porous anodic alumina: The formation of single nanopore and nanopore arrays

Anodized aluminium oxide (AAO) fabricated by electrochemical anodization of aluminium is used as the substrate for focused ion-beam (FIB) fabrication of single nanopore and nanopore arrays. The method is based on the controlled and selective removal of AAO barrier oxide film by FIB milling. FIB process with different milling conditions is applied to open single nanopores (diameter of 60 nm) or circular nanopore arrays with a different number of opened pores on AAO substrate. This process combines a low cost AAO substrate and FIB milling, and offers an economically viable alternative for the fabrication of nanopore devices with controlled dimensions for a range of applications in nanofluidics, molecular separations and biosensing.     

Fabrication of silicon dioxide submicron channels without nanolithography for single biomolecule detection

We successfully fabricated nanochannel arrays with silicon dioxide (SiO(2)) surfaces for single biomolecule detection. The SiO(2) nanochannel fabrication is based on the combination of anisotropic etching by potassium hydroxide (KOH) solution, local oxidation of silicon (LOCOS) and plasma etching of silicon. This fabrication technique is easily controllable and is a simple and practical solution for low-cost and high-throughput fabrication of nanofluidic channels. Thus, this technique enables the generation of nanochannels with various nanoscale dimensions without using nanolithography.     

Fabrication and characterization of nanopore-based electrodes with radii down to 2 nm

We report on the fabrication and characterization of gold nanoelectrodes with carefully controlled nanometer dimensions in a matrix of insulating silicon nitride. A focused electron beam was employed to drill nanopores in a thin silicon nitride membrane. The size and shape of the nanopores were studied with high-resolution transmission electron microscopy and electron-energy-loss two-dimensional maps. The pores were subsequently filled with gold, yielding conically shaped nanoelectrodes. The nanoelectrodes were examined by atomic and electrostatic force microscopy. Their applicability in electrochemistry was demonstrated by steady-state cyclic voltammetry. Pores with a radii down to 0.4 nm and electrodes with radii down to 2 nm are demonstrated.     

Electrochemical protection of thin film electrodes in solid state nanopores

Solid state nanopores are a core element of next-generation single molecule tools in the field of nano-biotechnology. Thin film electrodes integrated into a pore can interact with charges and fields within the pore. In order to keep the nanopore open and thus functional electrochemically induced surface alteration of electrode surfaces and bubble formation inside the pore have to be eliminated. This paper provides electrochemical analyses of nanopores drilled into TiN membranes which in turn were employed as thin film electrodes. We studied physical pore integrity and the occurrence of water decomposition yielding bubble formation inside pores by applying voltages between -4.5 and +4.5 V to membranes in various protection stages continuously for up to 24 h. During potential application pores were exposed to selected electrolyte-solvent systems. We have investigated and successfully eliminated electrochemical pore oxidation and reduction as well as water decomposition inside nanopores of various diameters ranging from 3.5 to 25 nm in 50 nm thick TiN membranes by passivating the nanopores with a plasma-oxidized layer and using a 90% solution of glycerol in water as KCl solvent. Nanopore ionic conductances were measured before and after voltage application in order to test for changes in pore diameter due to electrochemical oxidation or reduction. TEM imaging was used to confirm these observations. While non-passivated pores were electrochemically oxidized, neither electrochemical oxidation nor reduction was observed for passivated pores. Bubble formation through water decomposition could be detected in non-passivated pores in KCl/water solutions but was not observed in 90% glycerol solutions. The use of a protective self-assembled monolayer of hexadecylphosphonic acid (HDPA) was also investigated.     

Morphology and damping characteristics of woodceramics

Woodceramics has attracted more and more attention as ecomaterial at low cost in recent years. The purpose of this paper is to investigate morphology, damping characteristics of woodceramics and relationship between them. The experimental results reveal that pores in woodceramics can be divided into macropores, micropores and nanopores according to their dimensional size. Macropores are the majority of interconnected pores while most micropores are closed pores. Based on formation process, nanopores are supposed to consist of isolated nanopores and dendritic nanopores. Woodceramics exhibits significant damping gains, especially at room temperature, higher strain amplitude and higher vibration frequency. The values of damping capacity increase with vibration frequency and strain amplitude, decrease with testing temperature. Macropores and some nanopores have little but micropores and some other nanopores have much contribution to damping properties of woodceramics. The possible damping mechanisms of woodceramics at normal temperature are defect damping and thermoelastic damping; at high temperatures, interface damping is likely to responsible for the majority of the damping of woodceramics.     

Noninvasive Surface Coverage Determination of Chemically Modified Conical Nanopores that Rectify Ion Transport

Surface modification will change the surface charge density (SCD) at the signal-limiting region of nanochannel devices. By fitting the measured i-V curves in simulation via solving the Poisson and Nernst-Planck equations, the SCD and therefore the surface coverage can be noninvasively quantified. Amine terminated organosilanes are employed to chemically modify single conical nanopores. Determined by the protonation-deprotonation of the functional groups, the density and polarity of surface charges are adjusted by solution pH. The rectified current at high conductivity states is found to be proportional to the SCD near the nanopore orifice. This correlation allows the noninvasive determination of SCD and surface coverage of individual conical nanopores.