Obinaju, Blessing and Martin, Frank (2015) Application of attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy to measure sub-lethal effects of potential mutagens. PhD thesis, Lancaster University.
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Abstract
Techniques employed in vibrational spectroscopy monitor the vibrational modes of functional groups within biomolecules and enable a correlation between chemical information and histological structures. Interrogation of biological samples using infrared (IR) techniques generates spectrum with wavenumber-absorbance intensities specific to biomolecules within the sample. Methods are relatively non-destructive, and so samples can subsequently be analyzed by more conventional approaches. Analyses can be carried out ex vivo or in situ in living tissue, where a reference range of a designated normal state can be derived, and anything lying outside this range is potentially atypical. Computational approaches allow one to minimize within-category confounding factors. The application of vibrational spectroscopy in contaminant biomonitoring is a welcome development which has enabled the investigation of realtime contaminant exposure effects in the tissues of sentinels. IR techniques such as attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy, was able to detect changes in various tissue samples exposed to varying levels of polycyclic aromatic hydrocarbons (PAHs). This technique discriminated between spatial and temporal variations in the interrogated tissues. Multivariate analysis was able to relate the alterations at various regions of the fingerprint, to PAH exposure and was able to detect PAH exposure in tissues from sites with no documented knowledge of contamination. ATR-FTIR detected PAH-induced changes in isolated nuclei of cultured cell populations in G0/G1 and S- phases of the cell cycle. Findings from the various projects affirm, that techniques involved in IR spectroscopy are highly sensitive to minimal changes in cell molecules. The ability to generate rapid results in real-time is valuable and the wide variety of sample types which can be interrogated using IR techniques makes it a suitable technique for environment biomonitoring.