Quantification of host–microbe interactions by automated fluorescence microscopy

We describe an automated fluorescence microscopy-based assay that quantifies the invasion of mammalian cells by intracellular pathogens. Pathogens associated with host cell surfaces, intracellular pathogens and mammalian cells are directly counted based on their specific fluorescent labeling. Such approach utilizes automated image acquisition and processing, and is thus ideally suited for high-throughput analyses. This method was validated using Listeria monocytogenes as a model intracellular pathogen.