The collection of cerebrospinal fluid (CSF) plays a pivotal role in the diagnosis of central nervous system diseases. Prior training in this invasive procedure is essential to minimize the risk of harming animals. Because of this risk, stress and anxiety can influence the learning process. Simulators can be used to teach and learn invasive procedures.The aim of this mixed-methods study was to validate a CSF collection simulator and investigate students' perceptions of the simulator as an educational tool.The quantitative approach validated the simulator for face and content validity, and students provided a general evaluation of the simulator using surveys.The simulator's construct validity was measured by means of a global rating scale. Students' perceptions were investigated qualitatively using semi-structured interviews. Experts (n = 13) confirmed the simulator's face and content validity. Students (n = 16) evaluated the simulator as supportive of their learning. Results for construct validity demonstrated higher global rating scores from experts than from students. The scores for procedural performance and procedural knowledge and flow showed significant differences (p ≤.05).Analysis of interviews with students (n = 10) revealed four main themes: emotions, learning process, evaluation of the model, and CSF collection procedure. In conclusion, this study validated the use of the CSF simulator as an educational tool that can help students overcome some of their anxiety in relation to performing an invasive procedure.