Background: In the dynamic field of medical sciences, pathology laboratories are crucial for diagnosing diseases and guiding treatment decisions. The safety of laboratory personnel is paramount due to the inherent health risks associated with handling infectious agents, chemical hazards, and physical injuries. This study assesses the current level of safety knowledge and practices among laboratory technicians at Shree Krishna Medical College, aiming to identify areas for improvement and evaluate the impact of training on safety compliance. Materials and Methods: A cross-sectional study was conducted among 60 laboratory technicians at Shree Krishna Medical College's pathology laboratory. A structured questionnaire collected data on demographics, knowledge of safety guidelines, adherence to safety practices, and perceptions of the safety culture. Statistical analysis involved descriptive statistics and tests for associations between demographic factors and safety knowledge or practices. Results: The majority of technicians demonstrated a high level of knowledge regarding personal protective equipment (91.7%) but showed gaps in chemical safety (66.7%). Adherence to safety practices was high, with 90% always wearing protective equipment. Barriers to safety compliance included lack of time (33.3%) and inadequate training (25%). Training significantly improved safety knowledge (91.7% reported improvement) and adherence to practices (91.7% reported improvement). The study also highlighted a positive safety culture within the laboratory, though improvements in resource allocation and management support were suggested.Conclusion: Laboratory technicians at Shree Krishna Medical College exhibit a strong foundation in safety knowledge and practices, with training significantly enhancing safety compliance. Addressing identified barriers, such as time constraints and training gaps, is essential for fostering an even safer laboratory environment. Continuous professional development and strategic resource allocation are recommended to sustain and improve the safety culture.