PURPOSES : The aim of this study was to compare the performance of the QUEENSOD method and the gravity model in estimating Origin-Destination (O/D) tables for a large-sized microscopic traffic simulation network. METHODS : In this study, an expressway network was simulated using the microscopic traffic simulation model, VISSIM. The gravity model and QUEENSOD method were used to estimate the O/D pairs between internal and between external zones. RESULTS: After obtaining estimations of the O/D table by using both the gravity model and the QUEENSOD method, the value of the root mean square error (RMSE) for O/D pairs between internal zones were compared. For the gravity model and the QUEENSOD method, the RMSE obtained were 386.0 and 241.2, respectively. The O/D tables estimated using both methods were then entered into the VISSIM networks and calibrated with measured travel time. The resulting estimated travel times were then compared. For the gravity model and the QUEENSOD method, the estimated travel times showed 1.16% and 0.45% deviation from the surveyed travel time, respectively. CONCLUSIONS : In building a large-sized microscopic traffic simulation network, an O/D matrix is essential in order to produce reliable analysis results. When link counts from diverse ITS facilities are available, the QUEENSOD method outperforms the gravity model.