AbstractWhite-tailed deer (Odocoileus virginianus) are a main host for the adult life stages of tick species of medical and veterinary importance. Since white-tailed deer play a vital role in tick ecology, research has been conducted to understand this tick–host relationship. To date, research involving captive white-tailed deer and artificial infestation of these animals with ticks has focused on host suitability, the role of white-tailed deer in tick-borne diseases, and anti-tick vaccine research. The methodology reported for these studies was at times not descriptive and inconsistent regarding how and what region of the white-tailed deer was infested with ticks. Here, we propose a standardized method to artificially infest captive white-tailed deer with ticks for research purposes. The protocol describes a method proven effective to experimentally infest captive white-tailed deer with blacklegged ticks (Ixodes scapularis) to study tick–host interactions. The methods can be reliably transferred for experimental infestation of white-tailed deer by other multi-host and one-host tick species.