Single pores have found application in detecting and characterizing individual objects such as cells, particles, and even individual molecules. The experimental approach, called resistive-pulse technique, is often performed at symmetric electrolyte conditions so that the properties of the passing object remain constant in the course of measurement and translocation. Here we report experiments with highly charged mesoparticles passing through pores placed in contact with a pH gradient and demonstrate that this setup allows probing protonation and deprotonation of the particles. On the basis of fast diffusion of protons and submillisecond deprotonation/protonation kinetics of carboxyl groups, we expected that the particles would change their ionization state within a few milliseconds. However, our results show that the kinetics of protonation and deprotonation of the highly charged particles is significantly slower and exceeds 100 ms. We hypothesize that condensation of counterions that occurs on the particles at higher pH is responsible for the modified rates of protonation. The slowed-down deprotonation is attributed to modified local pH of the solution next to a highly charged surface. In addition, we show how electroosmotic flow of neutral particles through a pore in contact with pH gradient can probe modulations of local surface charge properties of the pore by voltage polarity.

Ionic transport at the nanoscale features phenomena that are not observed in larger systems. Nonlinear current–voltage curves characteristic of ionic diodes as well as ion selectivity are examples of effects observed at the nanoscale. Many man-made nanopore systems are inspired by biological channels in a cell membrane, thus measurements are often performed in aqueous solutions. Consequently, much less is known about ionic transport in nonaqueous systems, especially in solid-state electrolytes. Here we show ionic transport through single pores filled with gel electrolyte of poly(methyl methacrylate) (PMMA) doped with LiClO4 in propylene carbonate. The system has no liquid interface and the ionic transport occurs through the porous gel structure. We demonstrate that a conically shaped nanopore filled with the gel rectifies the current and works as a solid-state ionic diode.

Solid and gel electrolytes offer significant advantages for cycle stability and longevity in energy storage technologies. These advantages come with trade-offs such as reduced conductivity and ion mobility, which can impact power density in storage devices even at the nanoscale. Here we propose experiments aimed at exploring the ion transport properties of a hybrid electrolyte system of liquid and gel electrolytes with meso and nanoscale components. We focus on single pore systems featuring LiClO4-propylene carbonate and LiClO4-PMMA gel, which are model electrolytes for energy storage devices. We identified conditions at which the systems considered featured rectifying current–voltage curves, indicating a preferential direction of ion transport. The presented ion current rectification suggests different mechanisms arising from the unique hybrid system: (i) PMMA structure imposing selectivity in fully immersed systems and (ii) ionic selectivity linked to ion sourcing from media of different ionic mobility. These mechanisms were observed to interplay with ion transport properties linked to nanopore structure i.e. cylindrical and conical.

Conically shaped pores such as glass pipets as well as asymmetric pores in polymers became an important analytics tool used for the detection of molecules, viruses, and particles. Electrokinetic or pressure driven passage of single particles through a single pore causes a transient change of the transmembrane current, called a resistive-pulse, whose amplitude is the measure of the particle volume. The shape of the pulse reflects the pore topography, and in a conical pore, resistive pulses have a shape of a tick point. Passage of particles in both directions was reported to produce pulses of the same amplitude and shapes that are mirror images of each other. In this manuscript we identify conditions at which the amplitude of resistive-pulses in a conical mesopore is direction dependent. Neutral particles entering the pore from the larger entrance of a conical pore, called the base, block the current to a larger extent than the particles traveling in the opposite direction. Negatively charged particles on the other hand size larger when being transported in the direction from tip to base. The findings are explained via voltage-regulated ionic concentrations in the pore such that for one voltage polarity a weak depletion zone is formed, which increases the current blockage caused by a particle. For the opposite polarity, an enhancement of ionic concentrations was predicted. The findings reported here are of crucial importance for the resistive-pulse technique, which relates the current blockage with the size of the passing object.

Single pores in the resistive-pulse technique are used as an analytics tool to detect, size, and characterize physical as well as chemical properties of individual objects such as molecules and particles. Each object passing through a pore causes a transient change of the transmembrane current called a resistive pulse. In high salt concentrations when the pore diameter is significantly larger than the screening Debye length, it is assumed that the particle size and surface charge can be determined independently from the same experiment. In this article we challenge this assumption and show that highly charged hard spheres can cause a significant increase of the resistive-pulse amplitude compared to neutral particles of a similar diameter. As a result, resistive pulses overestimate the size of charged particles by even 20%. The observation is explained by the effect of concentration polarization created across particles in a pore, revealed by numerical modeling of ionic concentrations, ion current, and local electric fields. It is notable that in resistive-pulse experiments with cylindrical pores, concentration polarization was previously shown to influence ionic concentrations only at pore entrances; consequently, additional and transient modulation of resistive pulses was observed when a particle entered or left the pore. Here we postulate that concentration polarization can occur across transported particles at any particle position along the pore axis and affect the magnitude of the entire resistive pulse. Consequently, the recorded resistive pulses of highly charged particles reflect not only the particles’ volume but also the size of the depletion zone created in front of the moving particle. Moreover, the modeling identified that the effective surface charge density of particles depended not only on the density of functional groups on the particle but also on the capacitance of the Stern layer. The findings are of crucial importance for sizing particles and characterizing their surface charge properties.Keywords: concentration polarization; particles; resistive pulse

Biomimetic nanopores with rectifying properties are relevant components of ionic switches, ionic circuits, and biological sensors. Rectification indicates that currents for voltages of one polarity are higher than currents for voltages of the opposite polarity. Ion current rectification requires the presence of surface charges on the pore walls, achieved either by the attachment of charged groups or in multielectrode systems by applying voltage to integrated gate electrodes. Here we present a simpler concept for introducing surface charges via polarization of a thin layer of Au present at one entrance of a silicon nitride nanopore. In an electric field applied by two electrodes placed in bulk solution on both sides of the membrane, the Au layer polarizes such that excess positive charge locally concentrates at one end and negative charge concentrates at the other end. Consequently, a junction is formed between zones with enhanced anion and cation concentrations in the solution adjacent to the Au layer. This bipolar double layer together with enhanced cation concentration in a negatively charged silicon nitride nanopore leads to voltage-controlled surface-charge patterns and ion current rectification. The experimental findings are supported by numerical modeling that confirm modulation of ionic concentrations by the Au layer and ion current rectification even in low-aspect ratio nanopores. Our findings enable a new strategy for creating ionic circuits with diodes and transistors.

DNA strands can be analyzed at the single-molecule level by isolating them inside nanoscale holes. The strategy is used for the label-free and portable sequencing with nanopores. Nanochannels can also be applied to map genomes with high resolution, as shown by Jeffet et al. in this issue of ACS Nano. Here, we compare the two strategies in terms of biophysical similarities and differences and describe that both are complementary and can improve the DNA analysis for genomic research and diagnostics.

The ionic conductivity of porous MnO2 at the nanoscale is not well understood, despite possible importance in battery charging/discharging processes. It is demonstrated here that MnO2 in different oxidation states exhibits different ionic conductivities and surface charge characteristics, which are probed by reversal potential measurements.

Nanopores have become a model system to understand transport properties at the nanoscale. We report experiments and modeling of ionic current in aprotic solvents with different dipole moments through conically shaped nanopores in a polycarbonate film and through glass nanopipettes. We focus on solutions of the salt LiClO4, which is of great importance in modeling lithium based batteries. Results presented suggest ion current rectification observed results from two effects: (i) adsorption of Li+ ions to the pore walls, and (ii) a finite dipole moment rendered by adsorbed solvent molecules. Properties of surfaces in various solvents were probed by means of scanning ion conductance microscopy, which confirmed existence of an effectively positive surface potential in aprotic solvents with high dipole moments.

Single micropores in resistive-pulse technique were used to understand a complex dependence of particle mobility on its surface charge density. We show that the mobility of highly charged carboxylated particles decreases with the increase of the solution pH due to an interplay of three effects: (i) ion condensation, (ii) formation of an asymmetric electrical double layer around the particle, and (iii) electroosmotic flow induced by the charges on the pore walls and the particle surfaces. The results are important for applying resistive-pulse technique to determine surface charge density and zeta potential of the particles. The experiments also indicate the presence of condensed ions, which contribute to the measured current if a sufficiently high electric field is applied across the pore.

Hydrophobic nanopores provide a model system to study hydrophobic interactions at the nanoscale. Such nanopores could also function as a valve since they halt the transport of water and all dissolved species. It has recently been found that a hydrophobic pore can become wetted i.e. filled with condensed water or an aqueous solution of salt when a sufficiently high electric field is applied across the membrane. The wetting process is reversible thus when the voltage is lowered or switched off, the pore comes back to a closed state due to water evaporation in the pore. In this manuscript we present experimental studies on how the switching between conducting and non-conducting states can be regulated by the electrolyte concentration. Transport properties of single nanopores modified with alkyl chains of different lengths were recorded in salt concentrations between 10 mM and 1 M KCl. Nanopores modified with propyl chains exhibited gating in 10 mM KCl and were open for ionic transport for all voltages at higher salt concentrations. Nanopores modified with decyl chains did not conduct current in 10 mM and exhibited repeatable hydrophobic gating in 100 mM and 1 M KCl. The results are explained in the context of Maxwell stress in confined geometry with local surface charges, which change the shape of the water–vapor interface and promote wetting.

The resistive-pulse technique has been used to detect and size objects which pass through a single pore. The amplitude of the ion current change observed when a particle is in the pore is correlated with the particle volume. Up to date, however, the resistive-pulse approach has not been able to distinguish between objects of similar volume but different shapes. In this manuscript, we propose using pores with longitudinal irregularities as a sensitive tool capable of distinguishing spherical and rod-shaped particles with different lengths. The ion current modulations within resulting resistive pulses carry information on the length of passing objects. The performed experiments also indicate the rods rotate while translocating, and displace an effective volume that is larger than their geometrical volume, and which also depends on the pore diameter.Keywords: electroosmosis; excluded volume; pore; resistive-pulse; rod; rotation;

Biological channels embedded in cell membranes regulate ionic transport by responding to external stimuli such as pH, voltage, and molecular binding. Mimicking the gating properties of these biological structures would be instrumental in the preparation of smart membranes used in biosensing, drug delivery, and ionic circuit construction. Here we present a new concept for building synthetic nanopores that can simultaneously respond to pH and transmembrane potential changes. DNA oligomers containing protonatable A and C bases are attached at the narrow opening of an asymmetric nanopore. Lowering the pH to 5.5 causes the positively charged DNA molecules to bind to other strands with negative backbones, thereby creating an electrostatic mesh that closes the pore to unprecedentedly high resistances of several tens of gigaohms. At neutral pH values, voltage switching causes the isolated DNA strands to undergo nanomechanical movement, as seen by a reversible current modulation. We provide evidence that the pH-dependent reversible closing mechanism is robust and applicable for nanopores with opening diameters of up to 14 nm. The concept of creating an electrostatic mesh may also be applied to different organic polymers.

Pores with undulating opening diameters have emerged as an analytical tool enhancing the speed of resistive-pulse experiments, with a potential to simultaneously characterize size and mechanical properties of translocating objects. In this work, we present a detailed study of the characteristics of resistive-pulses of charged and uncharged polymer particles in pores with different aspect ratios and pore topography. Although no external pressure difference was applied, our experiments and modeling indicated the existence of local pressure drops, which modified axial and radial velocities of the solution. As a consequence of the complex velocity profiles, pores with undulating pore diameter and low-aspect ratio exhibited large dispersion of the translocation times. Distribution of the pulse amplitude, which is a measure of the object size, was not significantly affected by the pore topography. The importance of tuning pore geometry for the application in resistive-sensing and multipronged characterization of physical properties of translocating objects is discussed.

Rectifying nanopores feature ion currents that are higher for voltages of one polarity compared to the currents recorded for corresponding voltages of the opposite polarity. Rectification of nanopores has been found to depend on the pore opening diameter and distribution of surface charges on the pore walls as well as pore geometry. Very little is known, however, on the dependence of ionic rectification on the type of transported ions of the same charge. We performed experiments with single conically shaped nanopores in a polymer film and recorded current–voltage curves in three electrolytes: LiCl, NaCl, and KCl. Rectification degrees of the pores, quantified as the ratio of currents recorded for voltages of opposite polarities, were the highest for KCl and the lowest for LiCl. The experimental observations could not be explained by a continuum modeling based on the Poisson–Nernst–Planck equations. All-atom molecular dynamics simulations revealed differential binding between Li+, Na+, and K+ ions and carboxyl groups on the pore walls, resulting in changes to both the effective surface charge of the nanopore and cation mobility within the pore.

In this article we report resistive-pulse experiments with polystyrene particles whose transport through pores is controlled by modulating the driving voltage during the process of translocation. Balancing electric and hydrostatic forces acting on the particles allowed us to observe a random walk of single particles in a pore for tens of seconds and to quantify their diffusion coefficient using two methods. The first approach is based on the mean square displacement and requires passage of multiple particles for a range of diffusion times. The diffusion coefficient of individual particles was determined based on the variance of their local diffusion velocities. The developed methods for measuring the diffusion coefficient in pores are applicable to particles of different sizes, do not require fluorescence labeling, and are entirely based on ion current recordings. In addition, application of a modulating voltage signal together with rising edge triggers enabled transporting the same particle back and forth in the pore without letting the particle leave the pore.

We study the effect of electrolyte concentration on the shape of ion current pulses in resistive-pulse sensing. We show that electrokinetic passage of several hundred nanometers in diameter charged polystyrene particles through a micropore leads to formation of current increase when the particles exit the pore. The particle entrance, as reported before, causes formation of the current decrease, which is a measure of the particle size. Formation of the double peak, i.e., current decrease followed by a current increase, is especially pronounced if the resistive-pulse experiments are carried out in KCl concentrations below 200 mM. In order to explain the pulse shape, experiments were designed in which the particles passed through the pore only by either electroosmosis or electrophoresis. The presented experiments and modeling indicate that while both electroosmosis and electrophoresis affect the ion current pulse, formation of the positive peak is mainly determined by the latter effect and the charged state of the particle. The importance of the findings for resistive-pulse analysis is discussed.

The voltage-driven passage of biological polymers through nanoscale pores is an analytically, technologically, and biologically relevant process. Despite various studies on homopolymer translocation there are still several open questions on the fundamental aspects of pore transport. One of the most important unresolved issues revolves around the passage of biopolymers which vary in charge and volume along their sequence. Here we exploit an experimentally tunable system to disentangle and quantify electrostatic and steric factors. This new, fundamental framework facilitates the understanding of how complex biopolymers are transported through confined space and indicates how their translocation can be slowed down to enable future sensing methods.

Characterization of materials in confined spaces, rather than attempting to extrapolate from bulk material behavior, requires the development of new measurement techniques. In particular, measurements of individual meso- or nanoscale objects can provide information about their structure which is unavailable by other means. In this report, we perform measurements of ion currents through a few hundred nanometer long MnO2 rods deposited in single polymer pores. The recorded current confirms an existence of a meshlike character of the MnO2 structure and probes the effective size of the mesh voids and the polarity of surface charges. The recorded ion current through deposited MnO2 structure also suggests that the signal is mostly due to metal cations and not to protons. This is the first time that ionic current measurements have been used to characterize mesoporous structure of this important electrode material.

In this article, we report detection of deformable, hydrogel particles by the resistive-pulse technique using single pores in a polymer film. The hydrogels pass through the pores by electroosmosis and cause formation of a characteristic shape of resistive pulses indicating the particles underwent dehydration and deformation. These effects were explained via a non-homogeneous pressure distribution along the pore axis modeled by the coupled Poisson–Nernst–Planck and Navier–Stokes equations. The local pressure drops are induced by the electroosmotic fluid flow. Our experiments also revealed the importance of concentration polarization in the detection of hydrogels. Due to the negative charges as well as branched, low-density structure of the hydrogel particles, the concentration of ions in the particles is significantly higher than in the bulk. As a result, when an electric field is applied across the membrane, a depletion zone can be created in the vicinity of the particle observed as a transient drop of the current. Our experiments using pores with openings between 200 and 1600 nm indicated the concentration polarization dominated the hydrogels’ detection of pores wider than 450 nm. The results are of importance for all studies that involve transport of molecules, particles, and cells through pores with charged walls. The developed inhomogeneous pressure distribution can potentially influence the shape of the transported species. The concentration polarization changes the interpretation of the resistive pulses; the observed current change does not necessarily reflect only the particle size but also the size of the depletion zone that is formed in the particle vicinity.Keywords: deformation; hydrogels; pore; resistive-pulse technique

Hydrophobic interactions and local dewetting of hydrophobic cavities have been identified as a key mechanism for ionic gating in biological voltage-gated channels in a cell membrane. Hydrophobic interactions are responsible for rectification of the channels, i.e. the ability to transport ions more efficiently in one direction compared to the other. We designed single polymer nanopores with a hydrophobic gate on one side in the form of a single layer of C10 or C18 thiols. This nanoporous system behaves like an ionic diode whose direction of rectification is regulated by the pH of the electrolyte. In addition, reversible dewetting of the hydrophobic region of the pore was observed as voltage-dependent ion current fluctuations in time between conducting and non-conducting states. The observations are in accordance with earlier molecular dynamics simulations, which predicted the possibility of spontaneous and reversible dewetting of hydrophobic pores.

In this article, we report resistive-pulse sensing experiments with cylindrical track-etched PET pores, which reveal that the diameters of these pores fluctuate along their length. The resistive pulses generated by polymer spheres passing through these pores have a repeatable pattern of large variations corresponding to these diameter changes. We show that this pattern of variations enables the unambiguous resolution of multiple particles simultaneously in the pore, that it can detect transient sticking of particles within the pore, and that it can confirm whether any individual particle completely translocates the pore. We demonstrate that nonionic surfactant has a significant impact on particle velocity, with the velocity decreasing by an order of magnitude for a similar increase in surfactant concentration. We also show that these pores can differentiate by particle size and charge, and we explore the influence of electrophoresis, electroosmosis, and pore size on particle motion. These results have practical importance for increasing the speed of resistive-pulse sensing, optimizing the detection of specific analytes, and identifying particle shapes.Keywords: ion current; nanoparticle; nanopore; resistive-pulse technique

Ion currents through three types of rectifying nanoporous structures are studied and compared: conically shaped polymer nanopores, glass nanopipettes, and silicon nitride nanopores. Time signals of ion currents are analyzed by the power spectrum. We focus on the low-frequency range where the power spectrum magnitude scales with frequency, f, as 1/f. Glass nanopipettes and polymer nanopores exhibit nonequilibrium 1/f noise; thus, the normalized power spectrum depends on the voltage polarity and magnitude. In contrast, 1/f noise in rectifying silicon nitride nanopores is of equilibrium character. Various mechanisms underlying the voltage-dependent 1/f noise are explored and discussed, including intrinsic pore wall dynamics and formation of vortices and nonlinear flow patterns in the pore. Experimental data are supported by modeling of ion currents based on the coupled Poisson−Nernst−Planck and Navier−Stokes equations. We conclude that the voltage-dependent 1/f noise observed in polymer and glass asymmetric nanopores might result from high and asymmetric electric fields, inducing secondary effects in the pore, such as enhanced water dissociation.

Single nanopores attract a great deal of scientific interest as a basis for biosensors and as a system to study the interactions and behavior of molecules in a confined volume. Tuning the geometry and surface chemistry of nanopores helps create devices that control transport of ions and molecules in solution. Here, we present single conically shaped nanopores whose narrow opening of 8 or 12 nm is modified with single-stranded DNA molecules. We find that the DNA occludes the narrow opening of nanopores and that the blockade extent decreases with the ionic strength of the background electrolyte. The results are explained by the ionic strength dependence of the persistence length of DNA. At low KCl concentrations (10 mM) the molecules assume an extended and rigid conformation, thereby blocking the pore lumen and reducing the flow of ionic current to a greater extent than compacted DNA at high salt concentrations. Attaching flexible polymers to the pore walls hence creates a system with tunable opening diameters in order to regulate transport of both neutral and charged species.

The creation of synthetic devices that mimic functionality of biological systems is a task of fundamental importance for the future development of bio- and nanotechnology and also an ultimate test of our understanding of the biological systems. Among a plethora of bio-inspired devices, designed nanopores and nanochannels with an embedded functionality are of particular interest because of their potential applications in nanofluidic electronics, biosensing, separation, synthetic biology, and single-molecule manipulation. In this respect, nanopores with built-in stimulus-responsive properties are of special benefit. A transmembrane potential is a particularly useful stimulus as it is non-invasive, tunable, and can act over a short time scale. This critical review considers engineered solid-state and protein nanopores with voltage-responsive properties. The engineered systems show nonlinear current–voltage curves, and/or voltage-dependent switching between discrete conductance states (141 references).

Single conically shaped nanopores produce stable ion current fluctuations when in contact with weakly soluble salts, such as calcium hydrogen phosphate (CaHPO4) and cobalt hydrogen phosphate (CoHPO4). The pore spontaneously switches between high and low conductance states, called open and closed states, respectively. Pore opening and closing are linked to the dynamic formation of the calcium and cobalt precipitates at the small opening of the pore. The probabilities of pore opening and closing are voltage-dependent, and this characteristic of ion current signal is known for biological voltage-gated channels. We show that new types of ion current fluctuations are obtained in conditions at which precipitates of CaHPO4 and CoHPO4 can form in the pore at the same time.

Room temperature ionic liquids (RTILs) are substances composed entirely of ions and are liquids at or below 100 °C. Ionic conductivity of RTIL is one of the most important physical properties of these unique substances that determine their potential applications as a new medium for capacitors, fuel and solar cells as well as in separation systems. The quality of performance of these devices relies on the understanding of ionic transport of RTIL on a nanoscale. In this letter, we use ionic current carried by RTILs in single nanopores as a probe for their nanoscale transport properties. We show that the conductivity of RTILs through nanopores is significantly less than corresponding bulk values. Our experiments allowed us to address the nature of the interaction of these confined RTILs with charged surfaces. Electrostatic interactions of RTILs with nanopores are the basis for the formation of ionic diodes rectifying transport of the constituent ions.

Recently reported nanofluidic diodes with highly nonlinear current−voltage characteristics offer a unique possibility to construct different biosensors. These sensors are based on local changes of the surface charge on walls of single conical nanopores induced by binding of an analyte. The analyte binding can be detected as a change of the ion-current rectification of single nanopores defined as a ratio of currents for voltages of one polarity, and currents for voltages of the opposite polarity. In this article, we provided both modeling and experimental studies of various biosensing routes based on monitoring changes of the rectification degree in nanofluidic diodes used as a biosensing platform. A prototype of a sensor for the capsular poly γ-d-glutamic acid (γDPGA) from Bacillus anthracis is presented. The nanopore used for the sensing was locally modified with the monoclonal antibody for γDPGA. The proof of principle of the rectification degree-based sensing was further shown by preparation of sensors for avidin and streptavidin. Our devices also allowed for determination of the isoelectric point of the minute amounts of proteins immobilized on the surface.

Control of ionic transport through nanoporous systems is a topic of scientific interest for the ability to create new devices that are applicable for ions and molecules in water solutions. We show the preparation of an ionic transistor based on single conical nanopores in polymer films with an insulated gold thin film “gate.” By changing the electric potential applied to the “gate,” the current through the device can be changed from the rectifying behavior of a typical conical nanopore to the almost linear behavior seen in cylindrical nanopores. The mechanism for this change in transport behavior is thought to be the enhancement of concentration polarization induced by the gate.

The ionic conductivity of porous MnO2 at the nanoscale is not well understood, despite possible importance in battery charging/discharging processes. It is demonstrated here that MnO2 in different oxidation states exhibits different ionic conductivities and surface charge characteristics, which are probed by reversal potential measurements.

The creation of synthetic devices that mimic functionality of biological systems is a task of fundamental importance for the future development of bio- and nanotechnology and also an ultimate test of our understanding of the biological systems. Among a plethora of bio-inspired devices, designed nanopores and nanochannels with an embedded functionality are of particular interest because of their potential applications in nanofluidic electronics, biosensing, separation, synthetic biology, and single-molecule manipulation. In this respect, nanopores with built-in stimulus-responsive properties are of special benefit. A transmembrane potential is a particularly useful stimulus as it is non-invasive, tunable, and can act over a short time scale. This critical review considers engineered solid-state and protein nanopores with voltage-responsive properties. The engineered systems show nonlinear current–voltage curves, and/or voltage-dependent switching between discrete conductance states (141 references).