Automated image acquisition and analysis of stably expressing GFP-WIPI-1 U2OS cells with mCherry-expressing Staphylococci. Fluorescence images (middle panel) were automatically acquired using different emission/excitation filters for DAPI, GFP, and mCherry (left panel). DAPI and GFP images were used to automatically detect individual cells, and GFP images were used for detecting and analyzing GFP-WIPI-1 puncta formation (indicated in the right panel). Additionally, for each individual cell the bacterial area was determined (indicated in the right panel) and a fused image was further used to determine the number of cells harboring WIPI-1 positive autophagosome-like vesicles entrapping Staphylococci.
