A novel ex vivo set-up for dynamic long-term characterization of processes on mucosal interfaces by confocal imaging...

We present a novel organ-explant imaging system for easy and cost-effective extended-time observation of host-pathogen interactions at mucosal interfaces. Data are complemented by parallel cytokine measurements at high temporal resolution. The set-up is based on a custom-built reusable organ chamber compatible with standard microscopes. Luminal and basal side of the explanted mucosa are connected to separate channels for optimized incubation and cytokine measurements, oxygen is provided via membrane oxygenation. Dynamic imaging with confocal microscopy permits a detailed analysis of the dynamics of pathogen-host cell interactions at the mucosal interface and the neighbouring tissue at high resolution. The system can be applied to various hollow organs with few modifications. Here we present first applications to study representative infections such as uropathogenic Escherichia coli (UPEC) infections in the urinary bladder or amoebiasis of the colon by using mouse organs. We show (i) intracellular bacteria in UPEC infections, (ii) phagocytic events on tissue during infection, as well as (iii) tissue invasion of virulent protozoans into epithelia. The versatility of this system and its higher degree of control in comparison with both traditional explant microscopy and in vivo two photon imaging solutions make it a valuable and easy-to-use addition to other current imaging techniques.