Nanoparticle Charge and Shape Measurements using Tuneable Resistive Pulse Sensing

<p>Accurate characterisation of micro- and nanoparticles is of key importance in a variety of scientific fields from colloidal chemistry to medicine. Tuneable resistive pulse sensing (TRPS) has been shown to be effective in determining the size and concentration of nanoparticles in solution. Detection is achieved using the Coulter principle, in which each particle passing through a pore in an insulating membrane generates a resistive pulse in the ionic current passing through the pore. The distinctive feature of TRPS relative to other RPS systems is that the membrane material is thermoplastic polyurethane, which can be actuated on macroscopic scales in order to tune the pore geometry. In this thesis we attempt to extend existing TRPS techniques to enable the characterisation of nanoparticle charge and shape. For the prediction of resistive pulses produced in a conical pore we characterise the electrolyte solutions, pore geometry and pore zeta-potential and use known volume calibration particles. The first major investigation used TRPS to quantitatively measure the zeta-potential of carboxylate polystyrene particles in solution. We find that zeta-potential measurements made using pulse full width half maximum data are more reproducible than those from pulse rate data. We show that particle zeta-potentials produced using TRPS are consistent with literature and those measured using dynamic light scattering techniques. The next major task was investigating the relationship between pulse shape and particle shape. TRPS was used to compare PEGylated gold nanorods with spherical carboxylate polystyrene particles. We determine common levels of variation across the metrics of pulse magnitude, duration and pulse asymmetry. The rise and fall gradients of resistive pulses may enable differentiation of spherical and non-spherical particles. Finally, using the metrics and techniques developed during charge and shape investigations, TRPS was applied to Rattus rattus red blood cells, Shewanella marintestina bacteria and bacterially-produced polyhydroxyalkanoate particles. We find that TRPS is capable of producing accurate size distributions of all these particle sets, even though they represent nonspherical or highly disperse particle sets. TRPS produces particle volume measurements that are consistent with either literature values or electron microscopy measurements of the dominant species of these particle sets. We also find some evidence that TRPS is able to differentiate between spherical and non-spherical particles using pulse rise and fall gradients in Shewanella and Rattus rattus red blood cells. We expect TRPS to continue to find application in quantitative measurements across a variety of particles and applications in the future.</p>

Nanoparticle Charge and Shape Measurements using Tuneable Resistive Pulse Sensing

<p>Accurate characterisation of micro- and nanoparticles is of key importance in a variety of scientific fields from colloidal chemistry to medicine. Tuneable resistive pulse sensing (TRPS) has been shown to be effective in determining the size and concentration of nanoparticles in solution. Detection is achieved using the Coulter principle, in which each particle passing through a pore in an insulating membrane generates a resistive pulse in the ionic current passing through the pore. The distinctive feature of TRPS relative to other RPS systems is that the membrane material is thermoplastic polyurethane, which can be actuated on macroscopic scales in order to tune the pore geometry. In this thesis we attempt to extend existing TRPS techniques to enable the characterisation of nanoparticle charge and shape. For the prediction of resistive pulses produced in a conical pore we characterise the electrolyte solutions, pore geometry and pore zeta-potential and use known volume calibration particles. The first major investigation used TRPS to quantitatively measure the zeta-potential of carboxylate polystyrene particles in solution. We find that zeta-potential measurements made using pulse full width half maximum data are more reproducible than those from pulse rate data. We show that particle zeta-potentials produced using TRPS are consistent with literature and those measured using dynamic light scattering techniques. The next major task was investigating the relationship between pulse shape and particle shape. TRPS was used to compare PEGylated gold nanorods with spherical carboxylate polystyrene particles. We determine common levels of variation across the metrics of pulse magnitude, duration and pulse asymmetry. The rise and fall gradients of resistive pulses may enable differentiation of spherical and non-spherical particles. Finally, using the metrics and techniques developed during charge and shape investigations, TRPS was applied to Rattus rattus red blood cells, Shewanella marintestina bacteria and bacterially-produced polyhydroxyalkanoate particles. We find that TRPS is capable of producing accurate size distributions of all these particle sets, even though they represent nonspherical or highly disperse particle sets. TRPS produces particle volume measurements that are consistent with either literature values or electron microscopy measurements of the dominant species of these particle sets. We also find some evidence that TRPS is able to differentiate between spherical and non-spherical particles using pulse rise and fall gradients in Shewanella and Rattus rattus red blood cells. We expect TRPS to continue to find application in quantitative measurements across a variety of particles and applications in the future.</p>

Coordinated Detection of Micro- and Nanoparticles using Tuneable Resistive Pulse Sensing and Optical Spectroscopy

<p>The detection and characterisation of micro- and nanoscale particles has become increasingly important in many scientific fields, spanning from colloidal science to biomedical applications. Resistive Pulse Sensing (RPS) and its derivative Tuneable Resistive Pulse Sensing (TRPS), which both use the Coulter principle, have proven to be useful tools to detect and analyse particles in solution over a wide range of sizes. While RPS uses a fixed size pore, TRPS uses a dynamically stretchable pore in a polyurethane membrane, which has the advantages that the pore geometry can be tuned to increase the device's sensitivity and range of detection. The technique has been used to accurately determine the size, concentration and charge of many different analytes. However, the information obtained using TRPS does not give any insight into the particle's composition. In an attempt to overcome this, an experimental technique was developed in order to obtain simultaneous, time-resolved, high-resolution optical spectra of particles passing through the pore. Due to the ordered and controllable fashion in which the particles are guided through the sensing region, this approach has an advantage over diffusion based optical techniques. The experimental setup for the coordinated electrical and optical measurements involves many underlying physical phenomena, e.g. microuidics, electrokinetic effects, and Gaussian beam optics. A significant proportion of this work was therefore devoted to the development and the optimisation of the experimental setup by adapting a commercial TRPS device and a spectrometer with an attached microscope. Methods to engineer the spot size of a Gaussian beam to account for the different pore diameters, and the development of algorithms to filter, analyse and coordinate the recorded data are essential to the technique. The results using fluorescently labelled polystyrene particle sets with diameters from 190nm to 2 µm show that matching rates between the electrical and optical measurements of over 90% can repeatedly be achieved. Mixtures of particle species with similar diameters but with different fluorescent labels were used to demonstrate the technique's capability to characterise the analyte on a particle-by-particle basis and extend the information that can be obtained by TRPS alone. It was also shown that the data acquired with the electrical and optical measurements complement each other and can be used to better understand the TRPS technique itself. The influence of experimental parameters, such as the particle velocity, the beam size and the optical detection volume, on the intensity of the optical signals and the matching rates was studied intensively. These studies showed that the technique requires a careful experimental design to achieve the best results. Overall, the developed technique enhances the particle-by-particle specificity of conventional RPS measurements, and could be useful for a range of particle characterization and bio-analysis applications. Alongside the experiments, semi-analytic modelling and simulations using the Finite Element Method (FEM) were used to understand the particle motion through the pores, to interpret the experimental data, and predict the optical signals. The models were also used to assist the design and the optimization of the experiments. The FEM models were implemented with increasing physical detail and show superior understanding of the TRPS signals compared to the semi-analytic model, which is conventionally used in the TRPS field. The physical phenomena considered included o -axis trajectories, particle-field interactions for both fluid and electric fields, and the non-homogeneous distribution of ions close to the charged membrane and particle interfaces. Several effects which have been observed experimentally could be explained, including the intrinsic pulse height distribution, the current rectification, and the occurrence of bi-phasic pulses, demonstrating the benefits of FEM methods for RPS.</p>

Coordinated Detection of Micro- and Nanoparticles using Tuneable Resistive Pulse Sensing and Optical Spectroscopy

<p>The detection and characterisation of micro- and nanoscale particles has become increasingly important in many scientific fields, spanning from colloidal science to biomedical applications. Resistive Pulse Sensing (RPS) and its derivative Tuneable Resistive Pulse Sensing (TRPS), which both use the Coulter principle, have proven to be useful tools to detect and analyse particles in solution over a wide range of sizes. While RPS uses a fixed size pore, TRPS uses a dynamically stretchable pore in a polyurethane membrane, which has the advantages that the pore geometry can be tuned to increase the device's sensitivity and range of detection. The technique has been used to accurately determine the size, concentration and charge of many different analytes. However, the information obtained using TRPS does not give any insight into the particle's composition. In an attempt to overcome this, an experimental technique was developed in order to obtain simultaneous, time-resolved, high-resolution optical spectra of particles passing through the pore. Due to the ordered and controllable fashion in which the particles are guided through the sensing region, this approach has an advantage over diffusion based optical techniques. The experimental setup for the coordinated electrical and optical measurements involves many underlying physical phenomena, e.g. microuidics, electrokinetic effects, and Gaussian beam optics. A significant proportion of this work was therefore devoted to the development and the optimisation of the experimental setup by adapting a commercial TRPS device and a spectrometer with an attached microscope. Methods to engineer the spot size of a Gaussian beam to account for the different pore diameters, and the development of algorithms to filter, analyse and coordinate the recorded data are essential to the technique. The results using fluorescently labelled polystyrene particle sets with diameters from 190nm to 2 µm show that matching rates between the electrical and optical measurements of over 90% can repeatedly be achieved. Mixtures of particle species with similar diameters but with different fluorescent labels were used to demonstrate the technique's capability to characterise the analyte on a particle-by-particle basis and extend the information that can be obtained by TRPS alone. It was also shown that the data acquired with the electrical and optical measurements complement each other and can be used to better understand the TRPS technique itself. The influence of experimental parameters, such as the particle velocity, the beam size and the optical detection volume, on the intensity of the optical signals and the matching rates was studied intensively. These studies showed that the technique requires a careful experimental design to achieve the best results. Overall, the developed technique enhances the particle-by-particle specificity of conventional RPS measurements, and could be useful for a range of particle characterization and bio-analysis applications. Alongside the experiments, semi-analytic modelling and simulations using the Finite Element Method (FEM) were used to understand the particle motion through the pores, to interpret the experimental data, and predict the optical signals. The models were also used to assist the design and the optimization of the experiments. The FEM models were implemented with increasing physical detail and show superior understanding of the TRPS signals compared to the semi-analytic model, which is conventionally used in the TRPS field. The physical phenomena considered included o -axis trajectories, particle-field interactions for both fluid and electric fields, and the non-homogeneous distribution of ions close to the charged membrane and particle interfaces. Several effects which have been observed experimentally could be explained, including the intrinsic pulse height distribution, the current rectification, and the occurrence of bi-phasic pulses, demonstrating the benefits of FEM methods for RPS.</p>