Standardization is the trend of microfluidic system, therefore many devices require precise flow control and transport of fluid. Therefore, it’s important to characterize and measure the flow resistance for different chips. In this paper, a microfluidic analogue of the Wheatstone bridge circuit was developed for the measurement of different hydraulic resistance in a microfluidic device. It regards an on-chip membrane valve as a variable resistor, in parallel with the device. By applying different pressure, the membrane valve’s opening degree is controlled, Wheatstone bridge balance is kept, and the resistance of the chip is calculated directly. In this article, the measurement range reaching four magnitude was simulated on the computer with the error less than 3%. As a result, the microfluidic Wheatstone bridge represents an effective method for measurement of hydraulic resistance. And depending on various application environments, different dimensions and structures can be designed to satisfy special requirements.