Background <p>Technological Literacy (TL) is crucial in Biomedical Laboratory Science (BLS) for navigating the evolving Healthcare 4.0 landscape. This study explores TL’s conceptualization, operationalization, and implications for BLS education and practice.</p> Methods <p>Utilizing a mixed-methods approach, the study examined BLS curricula from University Colleges in Denmark. The research employed an explanatory sequential design, starting with quantitative data from questionnaires distributed to recent BLS graduates to identify trends and gaps. This was followed by qualitative interviews to gain deeper insights into these findings.</p> Results <p>The study finds a strong focus on application-oriented technology in BLS education. It identifies challenges such as curricular constraints, limited exposure to emerging trends, and the need for critical thinking skills. Recommendations include integrating critical thinking, ethical considerations, and experiential learning opportunities for supporting TL in BLS education. The research highlights the impact of hands-on experiences and authentic learning environments on student engagement with TL and stresses the importance of educator competencies, advocating for ongoing teacher training and professional development.</p> Conclusion <p>This study highlights the need for BLS programs to bridge theory and practice, emphasizing TL, hands-on learning, and faculty expertise. A comprehensive approach, integrating technical, ethical, and interdisciplinary skills, is essential to prepare graduates for a technology-driven healthcare landscape.</p> Clinical trial number <p>Not applicable.</p>