This work investigates the use of a fuzzy logic controller (FLC) for two-wheeled differential drive mobile robot trajectory tracking control. Due to the inherent complexity associated with tuning the membership functions of an FLC, this work employs a particle swarm optimization algorithm to optimize the parameters of these functions. In order to automate and reduce the number of rule bases, the genetic algorithm is also employed for this study. The effectiveness of the proposed approach is validated through MATLAB simulations involving diverse path tracking scenarios. The performance of the FLC is compared against established controllers, including minimum norm solution, closed-loop inverse kinematics, and Jacobian transpose-based controllers. The results demonstrate that the FLC offers accurate trajectory tracking with reduced root mean square error and controller effort. An experimental, hardware-based investigation is also performed for further verification of the proposed system. In addition, the simulation is conducted for various paths in the presence of noise in order to assess the proposed controller’s robustness. The proposed method is resilient against noise and disturbances, according to the simulation outcomes.