Biosensors using photonic crystal fibres

In this thesis, we present the unique properties of hollow-core photonic crystal fibres (HC-PCFs) for sensing applications in terms of viscosity detection and DNA sensing using a special poly(ethylene) glycol (PEGDA) hydrogel. The low loss HC-PCFs ensure a long interaction length between the sample and the optical signals. Thus in this thesis, we report the characterisation of filled HC-PCFs and the development of a selective filling process. For the first time, we report the investigation of a new viscometer device, and a new device for DNA sensing development, and also the chemical process for hydrogel growth was adapted to the fibres. By combining HC-PCFs with the hydrogel we enable 3D volumetric sample confinement within the HC-PCF, further increasing the interaction between the sample and the optical signal. However, the hydrogel has a large influence on the guidance properties of the HC-PCF and the HC-PCF has a strong influence on the growth process for the hydrogel itself. When we integrate the hydrogel and HC-PCFs we detect concentration levels as low as 400 nM of labelled DNA. However, using our technology for fluorescence detection we can achieve results two orders of magnitude better than those previously reported.

Gene delivery for the treatment of prostate cancer

Prostate cancer is one of the most common cancers diagnosed in men. Whilst treatments for early-stage disease are largely effective, current therapies for metastatic prostate cancer, particularly for bone metastasis, offer only a few months increased lifespan at best. Hence new treatments are urgently required. Small interfering RNA (siRNA) has been investigated for the treatment of prostate cancer where it can ‘silence’ specific cancer-related genes. However the clinical application of siRNA-based gene therapy is limited due to the absence of an optimised gene delivery vector. The optimisation of such gene delivery vectors is routinely undertaken in vitro using 2D cell culture on plastic dishes which does not accurately simulate the in vivo bone cancer metastasis microenvironment. The goal of this thesis was to assess the potential of two different targeted delivery vectors (gold or modified β-cyclodextrin derivatives) to facilitate siRNA receptor-mediated uptake into prostate cancer cells. Furthermore, this project aimed to develop a more physiologically relevant 3D in vitro cell culture model, to mimic prostate cancer bone metastasis, which is suitable for evaluating the delivery of nanoparticulate gene therapeutics. In the first instance, cationic derivatives of gold and β-cyclodextrin were synthesized to complex anionic siRNA. The delivery vectors were targeted to prostate cancer cells using the anisamide ligand which has high affinity for the sigma receptor that is overexpressed by prostate cancer cells. The gold nanoparticle demonstrated high levels of uptake into prostate cancer PC3 cells and efficient gene silencing when transfection was performed in serum-free media. However, due to the absence of a poly(ethylene glycol) (PEG) stabilising group, the formulation was unsuitable for use in serum-containing conditions. Conversely, the modified β-cyclodextrin formulation demonstrated enhanced stability in the presence of serum due to the inclusion of a PEG chain onto which the anisamide ligand was conjugated. However, the maximum level of gene silencing efficacy from three different prostate cancer cell lines (DU145, VCaP and PC3 cells) was 30 %, suggesting that further optimisation of the formulation would be required prior to application in vivo. In order to develop a more physiologically-relevant in vitro model of prostate cancer bone metastasis, prostate cancer cells (PC3 and LNCaP cells) were cultured in 3D on collagenbased scaffolds engineered to mimic the bone microenvironment. While the model was suitable for assessing nanoparticle-mediated gene knockdown, prostate cancer cells demonstrated a phenotype with lower invasive potential when grown on the scaffolds relative to standard 2D cell culture. Hence, prostate cancer cells (PC3 and LNCaP cells) were subsequently co-cultured with bone osteoblast cells (hFOB 1.19 cells) to enhance the physiological relevance of the model. Co-cultures secreted elevated levels of the MMP9 enzyme, a marker of prostate cancer metastasis, relative to prostate cancer cell monocultures (2D and 3D) indicating enhanced physiological relevance of the model. Furthermore, the coculture model proved suitable for investigating nanoparticle-mediated gene silencing. In conclusion, the work outlined in this thesis identified two different sigma receptor-targeted gene delivery vectors with potential for the treatment of prostate cancer. In addition, a more physiologically relevant model of prostate cancer bone metastasis was developed with the capacity to help optimise gene delivery vectors for the treatment of prostate cancer.

Formation of sub-7 nm feature size PS-b-P4VP block copolymer structures by solvent vapour process

The nanometer range structure produced by thin films of diblock copolymers makes them a great of interest as templates for the microelectronics industry. We investigated the effect of annealing solvents and/or mixture of the solvents in case of symmetric Poly (styrene-block-4vinylpyridine) (PS-b-P4VP) diblock copolymer to get the desired line patterns. In this paper, we used different molecular weights PS-b-P4VP to demonstrate the scalability of such high χ BCP system which requires precise fine-tuning of interfacial energies achieved by surface treatment and that improves the wetting property, ordering, and minimizes defect densities. Bare Silicon Substrates were also modified with polystyrene brush and ethylene glycol self-assembled monolayer in a simple quick reproducible way. Also, a novel and simple in situ hard mask technique was used to generate sub-7nm Iron oxide nanowires with a high aspect ratio on Silicon substrate, which can be used to develop silicon nanowires post pattern transfer.

Development of block copolymer lithography for device structure fabrication

This thesis investigated well-ordered block copolymer (BCP) thin film characteristics and their use for nanoscale pattern formation using a series of polystyrene-block-polymethylmethacrylate (PS-b-PMMA), polystyrene-blockpolydimethylsiloxane (PS-b-PDMS) and polystyrene-block-poly(ethylene oxide) (PS-b-PEO) systems of various molecular weights. BCP thin films, which act as an ‘on-chip’ etch mask and material templates, are highly promising self-assembling process for future scalable nanolithography. Unlike conventional BCP processing methods, the work in this thesis demonstrates that well-ordered patterns can be achieved in a few seconds compared to several hours by use of a non-conventional microwave assisted technique. As a result, well-ordered BCP nanoscale structures can be developed in industry appropriate periods facilitating their incorporation into current technologies. An optimised and controlled plasma dry etch process was used for successful pattern transfer to the underlying silicon substrate. Long range ordered BCP templates were further modified by selective metal inclusion technique to form a hard mask template towards fabrication of high aspect ratio nanopillars and nanowires. The work described here is centred on how these templates might be used to generate function at substrate surfaces. Herein we describe a number of innovations which might allow their successful uptake in a number of applications.