This study examined the challenges students face in learning Biology practical in public secondary schools in Dagoretti Sub-County, Nairobi County, Kenya. The research focused on the availability, accessibility, and frequency of use of Biology laboratory facilities, employing a descriptive survey design with a qualitative approach. Data was collected from a sample of students and Biology teachers to provide a comprehensive analysis of the factors affecting practical Biology education. Findings revealed that while most schools had Biology laboratories, many were shared with other subjects, limiting access and time allocation for Biology practical. Additionally, a significant number of students were unable to use the laboratory for independent study or group work outside scheduled class sessions. Teachers reported that time constraints, heavy workloads, and inadequate supervision further restricted the effective utilization of laboratory resources. Practical engagement was found to be infrequent, with many students rarely participating in hands-on activities, leading to a greater focus on theoretical learning rather than practical applications. The study concluded that despite the presence of laboratories, accessibility issues, resource shortages, and infrequent practical sessions negatively impacted students’ learning experiences. Inadequate laboratory equipment, lack of safety resources, and limited teacher availability hindered students from gaining essential practical skills. To address these challenges, the study recommended enhancing laboratory infrastructure, increasing resource allocation, scheduling more practical lessons, and adopting interactive learning approaches to improve student engagement. Strengthening laboratory management and ensuring policy implementation on practical learning were also suggested as ways to improve Biology practical education in public secondary schools.