Dielectrophoresis is a robust approach for the manipulation and separation of (bio)particles using microfluidic platforms. We developed a dielectrophoretic corral trap in a microfluidic device that utilizes negative dielectrophoresis to capture single spherical polystyrene particles. Circular‐shaped micron‐size traps were employed inside the device and the three‐dimensional trap stiffness (restoring trapping force from equilibrium trapping location) was analyzed using 4.42 μm particles and 1 MHz of an alternating electric field from 6 VP‐P to 10 VP‐P. The trap stiffness increased exponentially in the x‐ and y‐direction, and linearly in the z‐direction. Image analysis of the trapped particle movements revealed that the trap stiffness is increased 608.4, 539.3, and 79.7% by increasing the voltage from 6 VP‐P to 10 VP‐P in the x‐, y‐, and z‐direction, respectively. The trap stiffness calculated from a finite element simulation of the device confirmed the experimental results. This analysis provides important insights to predict the trapping location, strength of the trapping, and optimum geometry for single particle trapping and its applications such as single‐molecule analysis and drug discovery.