The blacklegged tick, Ixodes scapularis Say, is the primary vector of several tick-borne pathogens, including those causing Lyme disease and babesiosis, in the eastern United States and active collection methods for this species include dragging or wild animal sampling. Nest boxes targeting mice may be an alternative strategy for the surveillance and collection of immature I. scapularis feeding on these hosts and would be much safer for animals compared to small mammal trapping. We constructed double-walled insulated nest boxes (DWINs) with collection tubes mounted below the nesting chamber and deployed eleven in southern Wisconsin from June until September of 2020. The DWINs were occupied by Peromyscus spp. and birds (wren species, Troglodytidae family). We collected 192 ticks from collection tubes, all of which were identified as either I. scapularis (95%) or Dermacentor variabilis Say (Acari: Ixodidae) (5%). Only 12% (21/182) and 20% (2/10) of I. scapularis and D. variabilis were blood-fed, respectively. The high proportion of unfed ticks found in collection tubes may be due to grooming by hosts inside the nest boxes. Alternatively, immature ticks may have climbed trees and entered the DWIN seeking a host. Results suggest that nest boxes could be a tool for finding ticks in areas of low density or at the leading edge of invasion, when small mammal trapping or drag sampling is not feasible.