Since the NIH 'Sex as biological variable' policy, the percentage of studies including female subjects have increased largely. Nonetheless, many researchers fail to adequate their protocols to include females. In this narrative review, we aim to discuss the methodological pitfalls of the inclusion of female rodents in behavioral neuroscience. We address three points to consider in studies: the manipulations conducted only in female animals (such as estrous cycle monitoring, ovariectomy, and hormone replacement), the consideration of males as the standard, and biases related to interpretation and publication of the results. In addition, we suggest guidelines and perspectives for the inclusion of females in preclinical research.