This study aimed to evaluate tumour hypoxia by comparing [ 18 F]Fluoromisonidazole uptake measured using positron emission tomography ([ 18 F]FMISO-PET) with immunohistochemical (IHC) staining techniques. Syngeneic rhabdomyosarcoma (R1) tumour pieces were transplanted subcutaneously in the flanks of WAG/Rij rats. Tumours were analysed at volumes between 0.9 and 7.3 cm 3 . Hypoxic volumes were defined using a 3D region of interest on 2 h postinjection [ 18 F]FMISO-PET images, applying different thresholds (1.2 -3.0). Monoclonal antibodies to pimonidazole (PIMO) and carbonic anhydrase IX (CA IX), exogenous and endogenous markers of hypoxia, respectively, were used for IHC staining. Marker-positive fractions were microscopically measured for each tumour, and hypoxic volumes were calculated. A heterogeneous distribution of hypoxia was observed both with histology and
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