OBJECTIVE: Using a questionnaire, we wanted to gain insight into the knowledge of the laboratory information system by students of medical laboratory diagnostics and their perception of the importance of the laboratory information system. The goal was also to determine if there is a difference in the knowledge of the laboratory information system between students who have taken a course related to medical informatics and those who have not. STUDY DESIGN: Cross-sectional research conducted during August 2023. PARTICIPANTS AND METHODS: 44 respondents from the Undergraduate and Graduate Studies in Medical Laboratory Diagnostics in Osijek were included in the research. The research was conducted using a survey questionnaire that respondents filled out online. RESULTS: 40 girls and 4 boys participated in the survey. The median age of the respondents is 21 years. All students know about the term laboratory information system, but they have different levels of knowledge about it. Students who have taken a course related to medical informatics state significantly more that they are familiar with the program, subsystems and concept of the laboratory information system compared to those who have not. CONCLUSION: It can be concluded that students of Medical Laboratory Diagnostics are familiar with the laboratory information system and consider it an important and indispensable subsystem of the hospital information system. However, the differences in the depth of knowledge of the same differ between students who took a course related to medical informatics and those who did not.