In this study, a wheeled mobile robot navigation toolbox for Matlab is presented. The toolbox includes algorithms for 3D map design, static and dynamic path planning, point stabilization, localization, gap detection and collision avoidance. One can use the toolbox as a test platform for developing custom mobile robot navigation algorithms. The toolbox allows users to insert/remove obstacles to/from the robot’s workspace, upload/save a customized map and configure simulation parameters such as robot size, virtual sensor position, Kalman filter parameters for localization, speed controller and collision avoidance settings. It is possible to simulate data from a virtual laser imaging detection and ranging (LIDAR) sensor providing a map of the mobile robot’s immediate surroundings.... Differential drive forward kinematic equations and extended Kalman filter (EKF) based localization scheme is used to determine where the robot will be located at each simulation step. The LIDAR data and the navigation process are visualized on the developed virtual reality interface. During the navigation of the robot, gap detection, dynamic path planning, collision avoidance and point stabilization procedures are implemented. Simulation results prove the efficacy of the algorithms implemented in the toolbox.