Tick Blood Feeding through an Artificial Membrane on a 3D-Printed Chamber in Ixodid Tick Research

This work presents a detailed protocol for a tick feeding system through an artificial silicone membrane utilizing a 3D-printed chamber created from high-resolution photopolymer resin. The method includes the design of the feeding chamber, preparation of the silicone membrane, assembly of the system, and optimized conditions for tick feeding. Ticks are one of the leading vectors of disease in the United States and worldwide. Tick research is essential for developing effective strategies to prevent and reduce the incidence of tick-borne illnesses. As obligate hematophagous ectoparasites, ticks must feed on blood to survive and thus transmit disease, making blood-feeding a fundamental process when investigating vector-host interactions, pathogen transmission, and the biology, physiology, and anatomical morphology of ticks. Tick feeding systems with a silicone membrane have been widely utilized and refined over the past several decades. However, many existing systems are tailored to a single tick species, life stage, or specific set of conditions. The tick feeding system described in this protocol has been successfully used to obtain both partially fed and fully engorged specimens from several of the most important tick vector species in the United States, including Amblyomma americanum, A. maculatum, Ixodes scapularis, and Rhipicephalus sanguineus adults. The system has shown success in initiating feeding of Dermacentor variabilis adults and nymphal A. americanum, with the highest feeding rates (>85%) observed in adult A. americanum. Consequently, the feeding chamber developed in this study is adaptable for use with a low volume of different blood, down to 100 µL, which is crucial in pathogen transmission and toxicological studies. The transparent lid with minimal height of the chamber facilitated the observation of ticks over tick tick-feeding duration.