Virtual laboratory simulations (VLs), particularly in Biology education, are gaining popularity. This study focused on assessing students’ and teachers’ perceptions of VLs, developed by Labster, in a first-year undergraduate cell biology course. The evaluation involved surveys and interviews with 352 respondents. Findings indicate that over 90% of students found VLs user-friendly and visually engaging. However, around 60% noted the need for significant technical resources, which could limit accessibility. Over 80% of students reported that VLs increased their motivation, autonomy, interest, and confidence. While only 72% viewed them as potential replacements for traditional labs, more than 90% preferred using VLs as preparatory material for practical sessions. Teachers generally agreed with these perceptions, emphasizing the need for technical support for effective use. They suggested that VLs are better as pre-lab activities rather than full replacements. Both students and teachers recognized VLs as beneficial for academic performance and learning attitude but noted limitations in technical support for home use. The consensus was that VLs are most effective as complementary material before in-person lab sessions.