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Title: The development of a novel method for the Raman spectroscopic non-destructive analysis of tissue engineered bone cell cultures
Author: Thompson, Rachel Louise
ISNI:       0000 0004 6350 4388
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2017
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Raman spectroscopy is a burgeoning area in biological research and is of particular interest in providing a way to analyse tissue engineered constructs. However, current Raman spectroscopic methods are often suboptimal for use in biological spectroscopy and new instrumentation methods may be useful. This work proposes a new spectrometer design using lens vibration lock-in amplified Raman spectroscopy, which uses the principle that modulating the position of the laser focal spot generates a signal which is suitable for lock-in amplification. The key aim of the design was to enable low irradiance Raman spectroscopy using a maximum laser power of 20 mW at 788 nm, to minimise the risk of causing cell damage. The application of lockin amplification to this signal allows for phase selective amplification, selectively amplifying the Raman signal above the background. The spectrometer design was tested and validated initially by producing spectra from silica and 3 forms of hydroxyapatite. Next, cultures of mesenchymal stromal cells cultured in proliferation and differentiation media, and prepared for scanning electron microscopy were analysed. The spectra collected demonstrated that the application of lock-in amplification improved spectral quality compared to spectral collection without lock-in amplification. Interestingly the spectral signals collected from the cell cultures were also stronger than those collected from silica and hydroxyapatite with similar collection settings, suggesting that the preparation of the cells for scanning electron microscopy generated a surface-enhanced Raman effect. Raman mapping was then performed across the proliferative cell culture, generating a map across the sample at a wavenumber shift of 2149.1 cm-1, which demonstrated improvement with lock-in amplification. Finally, this work attempted to apply lens vibration lock-in amplification to a commercial CCD compact spectrometer. This effort, while not completely successful, represents a positive step towards producing commercially viable lens vibration lock-in amplified Raman spectroscopy.
Supervisor: Hatton, Paul V. ; Varcoe, Benjamin T. H. ; Crawford, Aileen Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available