Multiphoton microscopy applied for real-time intravital imaging of bacterial infections in vivo

To understand the underlying mechanisms of bacterial infections, researchers have for long addressed the molecular interactions occurring when the bacterium interacts with host target cells. In these studies, primarily based on in vitro systems, molecular details have been revealed along with increased knowledge regarding the general infection process. With the recent advancements in in vivo imaging techniques, we are now in a position to bridge a transition from classical minimalistic in vitro approaches to allow infections to be studied in its native complexity-the live organ. Techniques such as multiphoton microscopy (MPM) allow cellular-level visualization of the dynamic infection process in real time within the living host. Studies in which all interplaying factors, such as the influences of the immune, lymphatic, and vascular systems can be accounted for, are likely to provide new insights to our current understanding of the infection process. MPM imaging becomes extra powerful when combined with advanced surgical procedure, allowing studies of the illusive early hours of infection. In this chapter, our intention is to provide a general view on how to design and carry out intravital imaging of a bacterial infection. While exemplifying this using a spatiotemporally well-controlled uropathogenic Escherichia coli (UPEC) infection in rat kidneys, we hope to provide the reader with general considerations that can be adapted to other bacterial infections in organs other than the kidney.