A normally-open thermopneumatic-actuated microvalve has been fabricated and its properties are investigated. The advantages of the proposed microvalve are the low-cost fabrication process and the transparent properties of the polydimethylsiloxane (PDMS) and indium–tin–oxide (ITO)-coated glass. The fabricated microvalves with in-channel configuration are easily integrated with other microfluidic devices on the same substrate. The fabrication process of thermopneumatic-actuated microvalves using PDMS is simple and its performance is suitable for a disposable lab-on-a-chip. The deflection of the PDMS membrane increases and the flow rate of the microchannel and microvalve decreases as the applied power to the ITO heater increases. The power to cut off flow is dependent on the membrane thickness and the applied inlet pressure but is independent of the channel width of the microvalves. The flow rate could be easily controlled by the switching function of the ITO heater and the closing and opening times were around 20 and 25 s, respectively.