Biological vibrational spectroscopy: From a cellular to a sub-cellular level

The prevention of fatality from disease has long been a goal of medical and food sciences. In the field of cervical cancer research, the early detection of cancerous cells via Pap smear screening has proven to be an effective tool in combating cancer related deaths. However the sheer magnitude of Pap smears needing diagnosis has led to a decrease in efficiency and accuracy. In a different field, a similar problem persists. The goal of food science is to provide mass quantities of food that is safe for consumption. The amount of food prepared for dispersal is staggering in that it has prevented effective quality control of pathogenic contamination. Infrared (IR) spectroscopy has been shown to be useful in the diagnosis of cervical cancer. In this study a novel use of near-IR spectroscopy has been employed to assess its ability to distinguish between cancer and non-cancer without interfering with the current cervical cell preparation. In addition the introduction of a novel data visualization method enhances the difference between the cancerous and non-cancerous cells. NIR spectroscopy was found to be useful for accurate diagnosis of cervical cancer and easily introducable into the current detection system. Infrared (IR) spectroscopy has also been shown to be useful in the detection and classification of food borne pathogens. A novel method of IR spectroscopy referred to as hyperspectral imaging has recently been developed for general use. However, the application of hyperspectral imaging as a tool for quality control measures has not been explored. In this study, the use of mid-infrared hyperspectral imaging was assessed for feasibility for the detection and classification of pathogenic and non-pathogenic bacteria on food matrix and was found promising. However capable IR spectroscopy is, it has not been widely applied to disease and infection prevention and detection. One of the primary reasons is that the spectral signatures which are used for pathogen and cancer detection are not easily explained or even understood. To overcome this difficulty further study is required into the complex nature of the chemistry of biological organisms. In this study two novel methods were explored as a means of studying biology at the subcellular level. The first involves lysing bacteria cells and separating the cells with a liquid chromatograph coupled to an online infrared spectrometer (LCIR). This system allows for simultaneous purification and study of cellular components. Second, the complexity of even pure cellular components such as proteins is still too great to fully understand the structural significance of their spectra. Therefore, a novel method was accessed as to its ability to ascertain the spectra-structura relationships of peptides. This method is called generalized 2D correlation spectroscopy and used spectra from Raman, mid-infrared, and near-infrared measurements of a series of peptides