A Single-Entity Method for Actively Controlled Nucleation and High-Quality Protein Crystal Synthesis.

Lack of controls and understanding in nucleation, which proceeds crystal growth and other phase transitions, has been a bottleneck challenge in chemistry, materials, biology, and other fields. The exemplary needs for better methods for biomacromolecule crystallization include (1) synthesizing crystals for high-resolution structure determinations in fundamental research and (2) tuning the crystal habit and thus the corresponding properties in materials and pharmaceutical applications. Herein, a deterministic method is established capable of sustaining the nucleation and growth of a single crystal using the protein lysozyme as a prototype. The supersaturation is localized at the interface between a sample and a precipitant solution, spatially confined by the tip of a single nanopipette. The exchange of matter between the two solutions determines the supersaturation, which is controlled by electrokinetic ion transport driven by an external potential waveform. Nucleation and subsequent crystal growth disrupt the ionic current limited by the nanotip and are detected. The nucleation and growth of individual single crystals are measured in real time. Electroanalytical and optical signatures are elucidated as feedbacks with which active controls in crystal quality and method consistency are achieved: five out of five crystals diffract at a true atomic resolution of up to 1.2 Å. As controls, those synthesized under less optimized conditions diffract poorly. The crystal habits during the growth process are tuned successfully by adjusting the flux. The universal mechanism of nano-transport kinetics, together with the correlations of the diffraction quality and crystal habit with the crystallization control parameters, lay the foundation for the generalization to other materials systems.

Deconvolution of electroosmotic flow in hysteresis ion transport through single asymmetric nanopipettes.

Unveiling the contributions of electroosmotic flow (EOF) in the electrokinetic transport through structurally-defined nanoscale pores and channels is challenging but fundamentally significant because of the broad relevance of charge transport in energy conversion, desalination and analyte mixing, micro and nano-fluidics, single entity analysis, capillary electrophoresis etc. This report establishes a universal method to diagnose and deconvolute EOF in the nanoscale transport processes through current-potential measurements and analysis without simulation. By solving Poisson, Nernst-Planck (PNP) with and without Navier-Stokes (NS) equations, the impacts of EOF on the time-dependent ion transport through asymmetric nanopores are unequivocally revealed. A sigmoidal shape in the I-V curves indicate the EOF impacts which further deviate from the well-known non-linear rectified transport features. Two conductance signatures, an absolute change in conductance and a 'normalized' one relative to ion migration, are proposed as EOF impact (factor). The EOF impacts can be directly elucidated from current-potential experimental results from the two analytical parameters without simulation. The EOF impact is found more significant in intermediate ionic strength, and potential and pore size dependent. The less-intuitive ionic strength and size dependence is explained by the combined effects of electrostatic screening and non-homogeneous charge distribution/transport at nanoscale interface. The time-dependent conductivity and optical imaging experiments using single nanopipettes validate the proposed method which is applicable to other channel type nanodevices and membranes. The generalizable approach eliminates the need of simulation/fitting of specific experiments and offers previously inaccessible insights into the nanoscale EOF impacts under various experimental conditions for the improvement of separation, energy conversions, high spatial and temporal control in single entity sensing/manipulation, and other related applications.

Correlation of Ion Transport Hysteresis with the Nanogeometry and Surface Factors in Single Conical Nanopores.

Better understanding in the dynamics of ion transport through nanopores or nanochannels is important for sensing, nucleic acid sequencing and energy technology. In this paper, the intriguing nonzero cross point, resolved from the pinched hysteresis current-potential (i-V) curves in conical nanopore electrokinetic measurements, is quantitatively correlated to the surface and geometric properties by simulation studies. The analytical descriptions of the conductance and potential at the cross point are developed: the cross-point conductance includes both the surface and volumetric conductance; the cross-point potential represent the overall/averaged surface potential difference across the nanopore. The impacts by individual parameter such as pore radius, half cone angle, and surface charges are systematically studied in the simulation that would be convoluted and challenging in experiments. The elucidated correlation is supported by and offer predictive guidance for experimental studies. The results also offer more quantitative and systematic insights in the physical origins of the concentration polarization dynamics in addition to ionic current rectification inside conical nanopores and other asymmetric nanostructures. Overall, the cross point serves as a simple yet informative analytical parameter to analyze the electrokinetic transport through broadly defined nanopore-type devices.

History-dependent ion transport through conical nanopipettes and the implications in energy conversion dynamics at nanoscale interfaces.

The dynamics of ion transport at nanostructured substrate-solution interfaces play vital roles in high-density energy conversion, stochastic chemical sensing and biosensing, membrane separation, nanofluidics and fundamental nanoelectrochemistry. Further advancements in these applications require a fundamental understanding of ion transport at nanoscale interfaces. The understanding of the dynamic or transient transport, and the key physical process involved, is limited, which contrasts sharply with widely studied steady-state ion transport features at atomic and nanometer scale interfaces. Here we report striking time-dependent ion transport characteristics at nanoscale interfaces in current-potential (I-V) measurements and theoretical analyses. First, a unique non-zero I-V cross-point and pinched I-V curves are established as signatures to characterize the dynamics of ion transport through individual conical nanopipettes. Second, ion transport against a concentration gradient is regulated by applied and surface electrical fields. The concept of ion pumping or separation is demonstrated via the selective ion transport against concentration gradients through individual nanopipettes. Third, this dynamic ion transport process under a predefined salinity gradient is discussed in the context of nanoscale energy conversion in supercapacitor type charging-discharging, as well as chemical and electrical energy conversion. The analysis of the emerging current-potential features establishes the urgently needed physical foundation for energy conversion employing ordered nanostructures. The elucidated mechanism and established methodology can be generalized into broadly-defined nanoporous materials and devices for improved energy, separation and sensing applications.

Quantification of steady-state ion transport through single conical nanopores and a nonuniform distribution of surface charges.

Electrostatic interactions of mobile charges in solution with the fixed surface charges are known to strongly affect stochastic sensing and electrochemical energy conversion processes at nanodevices or devices with nanostructured interfaces. The key parameter to describe this interaction, surface charge density (SCD), is not directly accessible at nanometer scale and often extrapolated from ensemble values. In this report, the steady-state current-voltage (i-V) curves measured using single conical glass nanopores in different electrolyte solutions are fitted by solving Poisson and Nernst-Planck equations through finite element approach. Both high and low conductivity state currents of the rectified i-V curve are quantitatively fitted in simulation at less than 5% error. The overestimation of low conductivity state current using existing models is overcome by the introduction of an exponential SCD distribution inside the conical nanopore. A maximum SCD value at the pore orifice is determined from the fitting of the high conductivity state current, while the distribution length of the exponential SCD gradient is determined by fitting the low conductivity state current. Quantitative fitting of the rectified i-V responses and the efficacy of the proposed model are further validated by the comparison of electrolytes with different types of cations (K(+) and Li(+)). The gradient distribution of surface charges is proposed to be dependent on the local electric field distribution inside the conical nanopore.

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.

Transmembrane potential across single conical nanopores and resulting memristive and memcapacitive ion transport.

Memristive and memcapacitive behaviors are observed from ion transport through single conical nanopores in SiO(2) substrate. In i-V measurements, current is found to depend on not just the applied bias potential but also previous conditions in the transport-limiting region inside the nanopore (history-dependent, or memory effect). At different scan rates, a constant cross-point potential separates normal and negative hysteresis loops at low and high conductivity states, respectively. Memory effects are attributed to the finite mobility of ions as they redistribute within the negatively charged nanopore under applied potentials. A quantative correlation between the cross-point potential and electrolyte concentration is established.

Surface charge density determination of single conical nanopores based on normalized ion current rectification.

Current rectification is well known in ion transport through nanoscale pores and channel devices. The measured current is affected by both the geometry and fixed interfacial charges of the nanodevices. In this article, an interesting trend is observed in steady-state current-potential measurements using single conical nanopores. A threshold low-conductivity state is observed upon the dilution of electrolyte concentration. Correspondingly, the normalized current at positive bias potentials drastically increases and contributes to different degrees of rectification. This novel trend at opposite bias polarities is employed to differentiate the ion flux affected by the fixed charges at the substrate-solution interface (surface effect), with respect to the constant asymmetric geometry (volume effect). The surface charge density (SCD) of individual nanopores, an important physical parameter that is challenging to measure experimentally and is known to vary from one nanopore to another, is directly quantified by solving Poisson and Nernst-Planck equations in the simulation of the experimental results. The flux distribution inside the nanopore and the SCD of individual nanopores are reported. The respective diffusion and migration translocations are found to vary at different positions inside the nanopore. This knowledge is believed to be important for resistive pulse sensing applications because the detection signal is determined by the perturbation of the ion current by the analytes.