Alternative environments to real-life have been in recent demand in regard to lighting design and in museums. In this study, the effectiveness of the perception of the museum space in simulations or virtual-based environments is studied. Answers to a questionnaire regarding lighting in four different visual experiences are compared: Real-life, virtual-video-based, virtual-photo-based and virtual-render-based. A total of 117 participants were divided into four visual experience groups. Each group answered the same lighting related questions for four exhibition halls in the Natural History Museum of the University of Pisa (Italy), which is housed in the Monumental Charterhouse of Calci. The answers were analyzed using ANOVA and a T-test. The results show that virtual experiences can be acceptable alternatives to real-life experience as the answers were indifferent in more than half of the criteria, and no criterion was affected significantly by experience, regardless of the hall’s characteristics. However, it was found that the hall’s characteristics also had an impact on the perception of the criteria in different experiences. Controlled artificial lighting or uniformly distributed lighting (full day or artificial light) were found to be more indifferent to the experience.