The present study describes a technique to obtain consecutiveluteal samples by colpotomy. The animals received an epidural anesthesia and local anesthesia (vaginal vault) and after ten minutes the vaginal vault was incised with a scalpel blade and tissue was dissected to provide access to the pelvic cavity and to retract the ovary into the vagina. Then, a luteal biopsy was performed with a Yomann biopsy nipper. Signs indicative of pain and stress during the vaginal vault incision, traction of ovary or luteal biopsy were observed only in two collections. However, these signs were observed in ten collections during dissection of the vaginal wall and peritoneum. The occurrence of ataxia was observed in 26 collections and it was usually related to a longer duration of the procedure. Ataxia could be divided in light (15/26), moderate (6/26) and severe (5/26). The occurrence of ovarian adhesions ipsilateral to the incision was evaluated only in the initial four collections. Adhesions were present in 16 collections. The protocol described above provided a safe and efficient method to acquire luteal samples. The low incidence of adhesions allows the consecutive use of females without any interference in subsequent ovulations and collections.