A driver circuit with device variation compensation function and an operation method thereof are provided. The driver circuit includes a pull-up switch unit, an isolating switch and a pull-down switch unit. A first terminal of the pull-up switch unit is coupled to a first voltage. A second terminal of the pull-up switch unit is coupled to an output terminal of the driver circuit. A first terminal of the isolating switch is coupled to the second terminal of the pull-up switch unit. A first terminal of the pull-down switch unit is coupled to a second terminal of the isolating switch. A second terminal of the pull-down switch unit is coupled to a second voltage. The pull-down switch unit has a device variation compensation function.