This chapter discusses the field of diffuse fluorescence tomography in terms of fluorescence diffuse optical tomography (FDOT) and fluorescence molecular tomography (FMT). A brief overview of the forward photon propagation model is given. In addition, the techniques and challenges associated with solving the inverse problem, required for successful reconstruction, are discussed. Moreover, special attention is given to the different instrumentation used in diffuse fluorescence tomography. This includes the instrumentation associated with adequate illumination of the sample as well as efficient detection. Furthermore, the diverse applications of diffuse fluorescence tomography are explored, ranging from its use in biomarkers to preclinical applications and translational imaging. Finally, the chapter looks at the emerging technologies, which will shape the field in the near future.