Background: Evidence is lacking concerning a clear benefit of single-port laparoscopic cholecystectomy (SILC) and transvaginal cholecystectomy (TVC) over the classical laparoscopic cholecystectomy (CLC). In this study, we investigated the preferences of the operation techniques among female employees in a tertiary university clinic. Materials and Methods: Study participants in the department of general surgery and gynecology were interviewed regarding their personal felt preferences for the mentioned procedures using a standardized illustrated questionnaire. Results: A total of 111 participants were included in the study. In 70.3% of cases, the transvaginal approach was unknown. The classical techniques were preferred in 95.2% of respondents. Participants with a wish for children showed a higher preference for nontransvaginal techniques (P = .011). The acceptance rate of transvaginal techniques among employees of the department of gynecology was higher than those of the department of general surgery (P = .028). Conclusions: The overall acceptance rate for TVC is low. Especially in case of a wish for children, SILC and CLC represent the preferred techniques. The lack of popularity of TVC could be an explanation for the refusal of this technique. Among employees of the gynecologic department, a transvaginal approach was significantly more often accepted. The cosmetic outcome and the knowledge about an operation technique certainly influence the decision making for the preferred surgical method.