Oxidative stress and metabolic alterations derived from inflammation and tumour growth lead to hypoxia and angiogenesis in cancer and are associated with disease aggressiveness as well as the evolution of drug resistance. The interplay between blood oxygenation, tumour hypoxia and oxidative stress has yet to be fully understood. Furthermore, the interaction of these chemical processes with the physical microenvironment, including the extracellular matrix, appears to be important. There are few validated, non-invasive, methods to detect the spatiotemporal distribution of these processes and without such tools, their impact on tumour biology and cancer patient outcomes cannot be studied. To help elucidate this interplay, we develop and apply novel imaging methods to address this unmet need first in preclinical models then with a strong drive towards application in patients.