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Title: Non-invasive biochemical analysis of cells, tissues and tissue constructs with Raman micro-spectroscopy
Author: Swain, Robin
ISNI:       0000 0004 2737 5638
Awarding Body: Imperial College London
Current Institution: Imperial College London
Date of Award: 2008
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The high sensitivity and non-invasive sensing capabihties of Raman micro-spectroscopy have made this technique an attractive tool for biochemical analysis of cells and tissues. Biochemical characterization via interrogation of molecular vibrations offers several advantages over conventional histological, cellular, and molecular techniques, including rapid, non-invasive analysis in the absence of fixatives or labels, and compatibility with aqueous biological systems. Despite these advantages, the vast majority of biological Raman studies involve invasive sample processing (fixation, dehydration), which are liable to produce spectral artefacts. The focus of this thesis is specifically on the non-invasive analysis of live cell and tissue systems using Raman micro-spectroscopy. The Raman spectrum of a cell or tissue is a biochemical 'fingerprint', containing molecularlevel information about all cellular and tissue matrix biopolymers. Raman spectral signatures have previously been used to characterize cellular phenotype, tissue composition and architecture, and to monitor cellular processes and tissue development. This thesis demonstrates the novel application of Raman micro-spectroscopy to explore: • Biochemical differences between cells synchronized in different phases of the cell cycle. • Characterisation of primary alveolar epithelial type II (ATII) pneumocyte differentiation in vitro. • Biochemical differences between primary ATII cells, cancerous A549 cells and transformed type I-like cells derived from primary ATII cells. • Mineralised bone nodule formation in vitro by several cell sources (embryonic and mesenchymal stem cells, and mature osteoblasts). Trends in mineralization were identified over a 28-day period for a detailed comparative study. In each investigation, spectroscopic results were correlated with those obtained using gold-standard biological techniques. Qualitative and quantitative chemometric techniques were used to process and analyse Raman spectral data, with strong emphasis on multivariate analysis techniques to ensure robust and objective data analysis. The work presented herein demonstrates the tremendous potential of Raman micro-spectroscopy for the non-invasive analysis of live cell and tissue systems.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available