Intravital confocal microscopy of dermal innate immune responses to flea-transmitted Yersinia pestis

The technique known as intravital microscopy (IVM), when used in conjunction with transgenic mice expressing fluorescent proteins in various cell populations, is a powerful tool with the potential to provide new insights into host-pathogen interactions in infectious disease pathogenesis in vivo. Yersinia pestis, the causative agent of plague, is typically deposited in a host’s skin during feeding of an infected flea. IVM has been used to characterize the innate immune response to Y. pestis in the skin and identify differences between the responses to needle-inoculated and flea-transmitted bacteria that would have been difficult, if not impossible, to detect by other means. Here we describe techniques used to image the neutrophil response to flea-transmitted Y. pestis in the dermis of live mice using conventional confocal microscopy.