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Cultural heritage research and conservation practice seek to preserve our cultural heritage. Understanding the composition of the materials that comprise the artwork or cultural object is critical to inform collections management and preservation treatments; however methods of analysis are constrained to those that are either non-destructive or can obtain the desired information from micro-samples in order to retain the integrity of the object. Raman spectroscopy is an ideal technique for characterising cultural materials as it is non-destructive, requires relatively little sample preparation and utilises short measurement times. Micro-Raman is especially useful for examining micro-samples and painting cross sections due to its spatial resolution being sufficiently high to target individual pigment grains. Despite the non-destructive, micro-sample analysis advantages, there are limitations to the use of Raman spectroscopy in the cultural heritage conservation context. Firstly, fluorescence is frequently observed in cultural materials. Secondly, with the large number of compositionally complex and heterogenous materials encountered in conservation, there is a need for advanced methods that can deal with large sample datasets. These methods are needed to facilitate examination of both non-spatial data and spatial (imaging) data, to extract the maximum amount of information possible from the limited sample available. It is the aim of this thesis to demonstrate how Raman micro-spectroscopy can provide new and useful information about paintings using the following strategies. (1) Creation of a reliable pigment database supported by X-ray diffraction data to confirm the structural identity of each pigment. At the time this work was started, a spectral library for pigment identification was necessary to have a comprehensive set of pigment reference spectra against which to compare unknowns. A Raman spectral pigment reference library was developed comprising over 180 sample