Background: Analysis of unit cost is needed to calculate the cost of each unit in departments to determine the cost of various services in hospital. Hospital is an institution that provides healthcare service comprehensively in various departments, one of them being the laboratory unit. Routine hematology examination is one of most frequently enquired test by physician, which in turn raises the cost spent by hospital. We would like to find the unit cost of routine hematology examination with Activity-based Costing (ABC) method, and to compare the result with the real cost that the hospital applies. Materials and Methods: This is a qualitative study with a case study design. Data were obtained from interviews, observations, documents and medical records. Analysis of the cost was conducted with ABC method, and then be compared with the real cost that the hospital applies. Results: The unit cost of routine hematology examination using ABC method is Rp. 90.147, while the real cost is Rp. 83.756. The difference is Rp. 6.391. Conclusion: The unit cost based on ABC method is higher than the real cost applied by the hospital. The ABC method gives more accurate cost because of the overhead costing is charged into each activity related to the routine hematology examination. This should be a concern to the hospital, giving them insight about cost containment process, which in turn may yield more profit. Keyword: unit cost, real cost, ABC method, routine hematology examination