Apparatus and methods of calibrating a microfluidic flow sensor, in which the flow of a fluid through a flow sensor is stopped and a first value is read from the flow sensor, then the fluid is pumped through the flow sensor sequentially at first and second selected rates, and readings from the flow sensor of the flow rate are taken for each of the rates. The readings are used in a polynomial equation to determine the actual flow rate, which is used to calibrate the sensor. The flow sensor can be connected to a computer programmed to perform the calibration method, determine the actual flow rate of the sensor, and make appropriate adjustments to the flow rate of a pump.