There is an increasing demand for label-free methods of cell analysis in fields such as cell biology, drug discovery, and medical diagnostics. Many optical techniques have been developed to this end, which typically use physical properties such as refractive index to better visualise cells without the need for fluorescent markers. However, if chemical information about the sample is desired then these techniques provide limited information. Fourier Transform Infrared (FTIR) spectroscopic imaging has emerged as a powerful analytical tool that has been used to study a wide variety of biological samples. By using a focal plane array (FPA) detector, thousands of spatially resolved infrared spectra can be recorded simultaneously, creating a chemical map of a sample. FTIR spectroscopic imaging is also a label-free and inherently non-destructive form of measurement. However, due to the high molar absorptivity of water, the use of FTIR spectroscopic imaging to study living cells remains challenging. As cells must be maintained in an aqueous environment, severe restrictions are placed on the maximum path lengths that can be used in infrared spectroscopic imaging experiments. The purpose of this research was to further develop the application of micro attenuated total reflection (ATR)-FTIR spectroscopic imaging to the study of live cells. Micro ATR-FTIR spectroscopic imaging avoids the path length restriction seen in transmission mode, as the path length is not dependent on the sample’s thickness. This means micro ATR-FTIR spectroscopic imaging can be done in bulk aqueous environments. A robust methodology to culture adherent cells directly on an ATR crystal surface was developed, and the extent to which subcellular features can be visualised was investigated. Multiple-reflection ATR spectroscopy of live cells is also presented. This thesis then presents for the first time the detection of an exogenous molecule within a living cell by FTIR spectroscopic imaging. The recently developed analysis method disrelation mapping was also applied to a spectroscopic imaging dataset of a single living cell. Evidence of a different distribution of molecular states of water, compared to that seen in bulk water, was detected in living cells.