Bioluminescent Tracking of Aspergillus Infection

The A. fumigatus isolate Af1161 was transformed with luciferase reporter genes with the gpdA promoter as previously described with minor modifications (Brock et al., 2008 (link); Ding et al., 2013 (link)). Spores were obtained and suspended in PBS. The activity was confirmed using microplate reader (Spectra Max M2) and IVIS Lumina XR system (PerkinElmer) with a spore suspension containing 1% luciferase reporter reagent (Promega). Infected mice were introduced intraperitoneally with 100 μl D-luciferin in PBS (33.3 mg/ml) 10 min before detection. Mice were anesthetized using 2.5% isoflurane with a constant flow and then placed in an IVIS Lumina XR system chamber. Scanning was performed on days 2, 4, and 6. The region was cropped in the chest area from each animal, and the total bioluminescence signal intensity of the lung region was extracted and analyzed using Living Image 4.2 (Caliper Life Sciences).