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An encoded microwell array for multi-image analysis of cells

Sayers, Edward John 2012. An encoded microwell array for multi-image analysis of cells. PhD Thesis, Cardiff University.
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Abstract

In cell populations there is a great deal of heterogeneity. Cells can be seen to react different to the same stimulus despite being the same cell type and under the same conditions. Single cell analysis of a population is, therefore, crucial to understanding the nature of these differences. Using different technologies to study a single cell can help a researcher gather more data on the nature of these differences. The first part of this study looked at the development of a microwell array as a technique to allow correlative microscopy of non-adherent cell types. Laser ablation was used to generate microwells in both glass and the silicone polymer, polydimethylsiloxane (PDMS). Different methods were tested to increase quality of the microwells with the use of a sacrificial layer proving successful. Microwells were determined to be suitable for both live cell imaging and scanning electron microscopy imaging with correlative microscopy of non-adherent cells demonstrated. A new microwell design to allow the tracking of a single well through different imaging steps including sectioning was devised. Surface modification was required to make PDMS more cytophilic and different methods were investigated in the next part of this thesis. More cytocompatible surfaces were produced for PDMS using silanisation of the surface to produce amine moieties proved the most successful. In the final part of the thesis the cell penetrating peptide (CPP) octaargine was investigated along with the pro-apoptotic peptides PAD and the Bcl-2 converter peptide. The blebbing effects of the two peptides were analysed using non-adherent cells in microwells and fixed cells on a normal surface. Apoptosis as means of cell death by these peptides was disputed and the necrotic blebbing caused by theses peptides investigated.

Item Type: Thesis (PhD)
Status: Unpublished
Schools: Engineering
Subjects: Q Science > QH Natural history > QH426 Genetics
T Technology > TA Engineering (General). Civil engineering (General)
Uncontrolled Keywords: Multi-image analysis; Correlative microscopy; Encoded microwell array; CLEM; Cell penetrating peptides
Date of First Compliant Deposit: 30 March 2016
Last Modified: 04 Jun 2017 04:04
URI: http://orca.cf.ac.uk/id/eprint/32306

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