Figure 3.

Step-by-step procedure on the approach to set up tick organ cultures (unpublished from Grabowski and Kissinger, in preparation). A 96 well plate is used to establish individual culture wells for individual salivary gland pairs. One pair from each female tick is placed into each well, where 2–5 separate ticks (n= 2–5) can be used per treatment group. Culture plates are setup before dissecting unfed and fed female I. scapularis ticks occurs. Once salivary glands are loaded, TBFV infection is completed first and then followed with dsRNA transfections. Experimental work using salivary gland cultures from unfed and fed female I. scapularis ticks has been recently established (Grabowski et al., 2019; Grabowski et al., 2018; Grabowski, Tsetsarkin, et al., 2017). TBFV, tick-borne flavivirus.