Carbon Nanotube Arrays for Intracellular Delivery and Biological Applications

Introducing nucleic acids into mammalian cells is a crucial step to elucidate biochemical pathways, modify gene expression in immortalized cells, primary cells, and stem cells, and intoduces new approaches for clinical diagnostics and therapeutics. Current gene transfer technologies, including lipofection, electroporation, and viral delivery, have enabled break-through advances in basic and translational science to enable derivation and programming of embryonic stem cells, advanced gene editing using CRISPR (Clustered regularly interspaced short palindromic repeats), and development of targeted anti-tumor therapy using chimeric antigen receptors in T-cells (CAR-T). Despite these successes, current transfection technologies are time consuming and limited by the inefficient introduction of test molecules into large populations of target cells, and the cytotoxicity of the techniques. Moreover, many cell types cannot be consistently transfected by lipofection or electroporation (stem cells, T-cells) and viral delivery has limitations to the size of experimental DNA that can be packaged. In this dissertation, a novel coverslip-like platform consisting of an array of aligned hollow carbon nanotubes (CNTs) embedded in a sacrificial template is developed that enhances gene transfer capabilities, including high efficiency, low toxicity, in an expanded range of target cells, with the potential to transfer mixed combinations of protein and nucleic acids. The CNT array devices are fabricated by a scalable template-based manufacturing method using commercially available membranes, eliminating the need for nano-assembly. High efficient transfection has been demonstrated by delivering various cargos (nanoparticles, dye and plasmid DNA) into populations of cells, achieving 85% efficiency of plasmid DNA delivery into immortalized cells. Moreover, the CNT-mediated transfection of stem cells shows 3 times higher efficiency compared to current lipofection methods. Evaluating the cell-CNT interaction elucidates the importance of the geometrical properties of CNT arrays (CNT exposed length and surface morphology) on transfection efficiency. The results indicate that densely-packed and shortly-exposed CNT arrays with planar surface will enhance gene delivery using this new platform. This technology offers a significant increase in efficiency and cell viability, along with the ease of use compared to current standard methods, which demonstrates its potential to accelerate the development of new cell models to study intractable diseases, decoding the signaling pathways, and drug discovery

A General Strategy to Make an On-Demand Library of Structurally and Functionally Diverse SERS Substrates

Surface-enhanced Raman spectroscopy (SERS) is a powerful technique for sensing molecules proximal to suitable coinage metal surfaces. The physical structure of the SERS-active metal layer and its support is a key design parameter inspiring considerable, and frequently specialized, efforts in substrate fabrication. The necessary gold film structure can arise from both the metallization process and the underlying support structure, and the structure of the support can deliver additional functions including analytical capabilities such as physical filtering. We used electroless plating as a general approach to create a library of SERS substrates: SERS-active gold films on a range of supports made from a variety of materials, made with a mixture of simple and complex fabrication histories, and offering a selection of structurally-derived functions. The result was that supports with existing functions had their capabilities enhanced by the addition of SERS sensing. Electroless plating thus offers a host of beneficial characteristics 3 for nanofabricating multifunctional SERS substrates, including: tolerance to substrate composition and form factor; low equipment overhead requirements; process chemistry flexibility—including compatibility with conventional top-down nanofabrication; and a lengthy history of commercial application as a simple metallization technique. We gold-plated standard nanofabrication-compatible silicon nitride support surfaces with planar and porous architectures, and with native and polymer-grafted surface chemistries. We used the same plating chemistry to form SERS-active gold films on cellulose fibers arrayed in commercial filter paper and formed into nanocellulose paper. In a functional sense, we used electroless plating to augment nanoporous filters, chromatography platforms, and nanofabrication building blocks with SERS capability

Development of Multifunctional E-skin Sensors

Electronic skin (e-skin) is a hot topic due to its enormous potential for health monitoring, functional prosthesis, robotics, and human-machine-interfaces (HMI). For these applications, pressure and temperature sensors and energy harvesters are essential. Their performance may be tuned by their films micro-structuring, either through expensive and time-consuming photolithography techniques or low-cost yet low-tunability approaches. This PhD thesis aimed to introduce and explore a new micro-structuring technique to the field of e-skin – laser engraving – to produce multifunctional e-skin devices able to sense pressure and temperature while being self-powered. This technique was employed to produce moulds for soft lithography, in a low-cost, fast, and highly customizable way. Several parameters of the technique were studied to evaluate their impact in the performance of the devices, such as moulds materials, laser power and speed, and design variables. Amongst the piezoresistive sensors produced, sensors suitable for blood pressure wave detection at the wrist [sensitivity of – 3.2 kPa-1 below 119 Pa, limit of detection (LOD) of 15 Pa], general health monitoring (sensitivity of 4.5 kPa-1 below 10 kPa, relaxation time of 1.4 ms, micro-structured film thickness of only 133 µm), and robotics and functional prosthesis (sensitivity of – 6.4 × 10-3 kPa-1 between 1.2 kPa and 100 kPa, stable output over 27 500 cycles) were obtained. Temperature sensors with micro-cones were achieved with a temperature coefficient of resistance (TCR) of 2.3 %/°C. Energy harvesters based on micro-structured composites of polydimethylsiloxane (PDMS) and zinc tin oxide (ZnSnO3) nanowires (NWs; 120 V and 13 µA at > 100 N) or zinc oxide (ZnO) nanorods (NRs; 6 V at 2.3 N) were produced as well. The work described herein unveils the tremendous potential of the laser engraving technique to produce different e-skin devices with adjustable performance to suit distinct applications, with a high benefit/cost ratio