Tick-borne viral infections, like those described in this book, pose real threats to the health of humans across the world. Powassan virus (POWV) and Deer tick virus (DTV) are two examples of tick-borne flaviviruses that are endemic to North America and cause neuroinvasive disease. These two viruses are both genetically similar and are spread by similar vectors but have differing tropism and clinical outcomes. Studies involving these viruses have routinely consisted of mouse models and in vitro studies. These models, effective as they are, are limited in their use as they are either costly or do not meet niche needs. Herein, we describe a method of using an ex vivo human skin model for infection studies that focus on the primary site of infection, the skin.