The use of Lewis rats in embryonic tissue transplantation experiments can present a challenge because of the fertility problems associated with this strain. The authors used estrous cycle phase determination to time pairings of reproductively active females and males in order to increase the likelihood that mating would occur. During a 24-month period, female rats in a production colony were evaluated for estrous phase by microscopic evaluation of vaginal smears and by vaginal impedance readings. Pairings that were arranged with females determined to be in the proestrous phase by vaginal smears resulted in a low rate of confirmed matings (14%). Serial vaginal smear sampling also produced a high incidence of pseudopregnancy. Pairings that were arranged with females determined to be in the proestrous phase by vaginal impedance readings resulted in a high rate of confirmed matings (48.1%) as well as a high rate of pregnancy (29.4%). An average of 7.2 embryos were produced per pregnancy. The vaginal impedance technique can be used to improve the breeding success rate of Lewis rats in order to maintain a stable production of embryos for experimentation.