Intravital Imaging of Vaccinia Virus-Infected Mice

Intravital multiphoton microscopy (MPM) allows the direct visualization of viral infections in real time as they occur in living animals. Here we describe the routes and considerations for murine infection with vaccinia virus (VACV) for imaging, and the preparation of the skin and inner lip (labial mucosa) of infected animals for MPM. Using different recombinant VACVs expressing fluorescent proteins in combination with transgenic fluorescent reporter mice, MPM imaging can be used to examine the movements, interactions, and functions of virus-infected cells or selected immune cell populations after infection.