During hypoxia, tissues are subjected to an inadequate oxygen supply, disrupting the balance needed to maintain normal function. This deficiency can occur due to reduced oxygen delivery caused by impaired blood flow or a decline in the blood's ability to carry oxygen. In tumors, hypoxia and vascularization play crucial roles, shaping their microenvironments and influencing cancer progression, response to treatment and metastatic potential. This chapter provides guidance on the use of non-invasive imaging methods including Positron Emission Tomography and Magnetic Resonance Imaging to study tumor oxygenation in pre-clinical settings. These imaging techniques offer valuable insights into tumor vascularity and oxygen levels, aiding in understanding tumor behavior and treatment effects. For example, PET imaging uses tracers such as [18F]-fluoromisonidazole (FMISO) to visualize hypoxic areas within tumors, while MRI complements this with anatomical and functional images. Although directly assessing tumor hypoxia with MRI remains challenging, techniques like Blood Oxygen Level Dependent (BOLD) and Dynamic Contrast-Enhanced MRI (DCE-MRI) provide valuable information. BOLD can track changes in oxygen levels during oxygen challenges, while DCE-MRI offers real-time access to perfusion and vessel permeability data. Integrating data from these imaging modalities can help assess oxygen supply, refine treatment strategies, enhance therapeutic effectiveness, and ultimately improve patient outcomes.

Centre for Nuclear Medicine Department of Radiology Haukeland University Hospital Bergen Norway; Molecular Imaging Center Department of Biomedicine University of Bergen Norway

Institute for Biomedical Research Sols Morreale Spanish National Research Council Universidad Autónoma de Madrid Madrid Spain

Institute of Scientific Instruments of the Czech Academy of Sciences Brno Czech Republic

Molecular Imaging Center Department of Biomedicine University of Bergen Norway

Molecular Imaging Center Department of Biomedicine University of Bergen Norway; Department of Neurosurgery Haukeland University Hospital Bergen Norway; Department of Neurosurgery Qilu Hospital of Shandong University and Brain Science Research Institute Shandong University Key Laboratory of Brain Functional Remodeling Shandong Jinan China

Molecular Imaging Center Department of Biomedicine University of Bergen Norway; Western Australia National Imaging Facility The University of Western Australia Perth Australia

Translational Radiomics Luxembourg Institute of Health Luxembourg City Luxembourg; In Vivo Imaging Platform Luxembourg Institute of Health Luxembourg City Luxembourg

Translational Radiomics Luxembourg Institute of Health Luxembourg City Luxembourg; In Vivo Imaging Platform Luxembourg Institute of Health Luxembourg City Luxembourg; Faculty of Electrical Engineering and Communication Brno University of Technology Brno Czech Republic

Tumor Immunotherapy and Microenvironment Group Department of Cancer Research Luxembourg Institute of Health Luxembourg City Luxembourg

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