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Current diagnostic methods for invasive aspergillosis are time-consuming and poorly sensitive. Here, the authors show that positron emission tomography with 2-deoxy-2-18F-fluorosorbitol can visualize Aspergillus fumigatus infection of the lungs, brain and muscles in mouse models, and can distinguish pulmonary aspergillosis from other diseases such as Staphylococcus aureus infection and lung cancer.