The use of a "size-tunable" polyurethane resistive pulse sensor for quantitative sizing of nano- and microparticles is presented. A linear relationship, as first suggested by Maxwell, between particle volume and change in electric resistance across the pore was observed. Particle sizes were quantified for a given size-tunable membrane, by first creating a linear calibration curve to a series of monodisperse carboxylated polystyrene particles of various diameters and then applying this curve to calculate the size of "unknown" nanoparticles. The diameters of a selection of synthetic and biological particles, being PMMA and nonfunctionalized polystyrene particles, along with biological nanoparticles (adenovirus) were calculated using this methodology. Calculated particle diameters and coefficients of variation were shown to be in good agreement with both transmission electron microscopy and dynamic light scattering results.
Tunable nanopores fabricated in elastomeric membranes have been used to study the dependence of ionic current blockade rate on the concentration and electrophoretic mobility of particles in aqueous suspensions. A range of nanoparticle sizes, materials and surface functionalities has been tested. Using pressure-driven flow through a pore, the blockade rate for 100 nm carboxylated polystyrene particles was found to be linearly proportional to both transmembrane pressure (between 0 and 1.8 kPa) and particle concentration (between 7 × 10(8) and 4.5 × 10(10) ml( - 1)). This result can be accurately modelled using Nernst-Planck transport theory, enabling measurement of particle concentrations. Using only an applied potential across a pore, the blockade rates for carboxylic acid and amine coated 500 and 200 nm silica particles were found to correspond to changes in their mobility as a function of the solution pH. Scanning electron microscopy and confocal microscopy have been used to visualize changes in the tunable nanopore geometry in three dimensions as a function of applied mechanical strain. The pores were conical in shape, and changes in pore size were consistent with ionic current measurements. A zone of inelastic deformation adjacent to the pore has been identified as important in the tuning process.
A novel method using resistive pulse sensors for electrokinetic surface charge measurements of nanoparticles is presented. This method involves recording the particle blockade rate while the pressure applied across a pore sensor is varied. This applied pressure acts in a direction which opposes transport due to the combination of electro-osmosis, electrophoresis, and inherent pressure. The blockade rate reaches a minimum when the velocity of nanoparticles in the vicinity of the pore approaches zero, and the forces on typical nanoparticles are in equilibrium. The pressure applied at this minimum rate can be used to calculate the zeta potential of the nanoparticles. The efficacy of this variable pressure method was demonstrated for a range of carboxylated 200 nm polystyrene nanoparticles with different surface charge densities. Results were of the same order as phase analysis light scattering (PALS) measurements. Unlike PALS results, the sequence of increasing zeta potential for different particle types agreed with conductometric titration.
Tunable resistive pulse sensing (TRPS) is an experimental technique that has been used to study and characterise colloidal particles ranging from approximately 50 nm in diameter up to the size of cells. The primary aim of this Review is to provide a guide to the characteristics and roles of TRPS in recent applied research. Relevant studies reflect both the maturation of the technique and the growing importance of submicron colloids in fields such as nanomedicine and biotechnology. TRPS analysis of extracellular vesicles is expanding particularly swiftly, while TRPS studies also extend to on-bead assays using DNA and aptamers, drug delivery particles, viruses and bacteria, food and beverages, and superparamagnetic beads. General protocols for TRPS measurement of particle size, concentration and charge have been developed, and a summary of TRPS technology and associated analysis techniques is included in this Review.
Asymmetric resistive pulses caused by nanoparticles passing through tunable nanopores have been recorded and studied using a semianalytic physical model. Experiments used 220 nm diameter carboxylate-modified polystyrene spheres, electrophoretically driven through two elastomeric nanopore specimens. Asymmetry is evident both within the pulse full-width half-maximum and over a longer 5 ms window. This asymmetry is consistent with the near-conical pore geometry, and is greater for both large and slow-moving particles. Particle mobility did not increase with size, and was unexpectedly enhanced when the electrolyte pH was reduced from 8.0 to 7.0. In the model, an on-axis insulating particle with an effective electrophoretic charge is suspended in an electrolyte of homogeneous resistivity. End effects, particle transport, and any azimuthally symmetric pore geometry are supported. When a linear cone geometry was fitted to experiments, values for the pore opening radii and the particle effective charge were obtained. More complicated geometries can better reproduce experimental pulse asymmetry and absolute sizes of pore openings. Nanopore-based resistive pulse measurement is being applied to sensing and analysis of many submicron particle types, including viruses, synthetic nanoparticles, and single molecules.
A way through the labyrinth: LabKC is an enzyme encoded in the biosynthesis gene cluster (lab gene cluster) of the labyrinthopeptin producer Actinomadura namibiensis. Some of its genes are homologous with those in other actinomycetes strains, for example, the model organism Streptomyces coelicolor. The functional assignment of LabKC as a kinase–cyclase suggests the formation of a new post‐translational modification by a consecutive double Michael addition (see scheme; Dha=α,β‐dehydroalanine).
Mechanical resizing of individual nanopores in a thermoplastic polyurethane elastomer has been characterized. Specimen nanopores were conical, with smaller hole dimensions of the order of tens to hundreds of nanometres. Electrophoretic current measurements show that the estimated nanopore radius can be reversibly actuated over an order of magnitude by stretching and relaxing the elastomer. Within a working range of stretching, current is proportional to specimen extension to the power of a constant, n, which ranges from 0.9 to 2.3 for different specimens. The data indicate that scaling of the effective pore radius is super-affine. At strains below the working range, the pore size is relatively unresponsive to stretching. Macroscopic elastomer extension has been related to local radial strain (50-250 µm from the pore) using optical microscopy. Scanning electron microscopy and atomic force microscopy have been used to observe membrane surface features.
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