Bioluminescence imaging (BLI) is widely used in pre-clinical research to monitor the location and migration of different cell types, and the growth of cancerous tumours and response to treatments within murine models. However, the quantitative accuracy of the technique is limited. The position of the animal is known to affect the measured bioluminescence, with a change in position causing a change in measurement. Work presented here will address this problem, validating a free space model in a murine model to produce surface bioluminescence measurements which are independent of the position of the animal. The position of the source within the animal and the underlying tissue attenuation also affect the quantitative accuracy of bioluminescence measurements. An extension to bioluminescence imaging, bioluminescence tomography (BLT), aims to overcome these problems by recovering the three-dimensional bioluminescent source distribution within the animal. However, there are limitations to the quantitative accuracy of BLT. Current reconstruction algorithms ignore the bandwidth of band-pass filters used for multi-spectral data collection for BLT. This work develops a model which accounts for filter bandwidth in the BLT reconstruction, improving the quantitative accuracy of the technique. An additional limitation to the quantitative accuracy of BLT is that accurate knowledge of the optical properties of the animal are required but are difficult to acquire. Work to improve the quantitative accuracy by obtaining subject-specific optical properties via a spectral derivative reconstruction method for diffuse optical tomography (DOT) is presented. The initial results are promising for the application of the method in vivo.