Fabrication of microchannels on polymethylmethacrylate (PMMA) substrates using novel microfabrication methods is demonstrated. The image of microchannels is transferred from a silicon master possessing the inverse image of the microchannel to a PMMA plate by using hot embossing methods. The silicon master is electrostatically bonded to a Pyrex 7740 glass wafer, which improves the device yield from about 20 devices to hundreds of devices per master. Effects of embossing temperature, pressure and time on the accuracy of replication are systematically studied using the orthogonal factorial design. According to the suggested experimental model, the time for the whole embossing procedure is shorten from about 20 min to 6 min, and the accuracy of replication is 99.3%. The reproducibility of the hot embossing method is evaluated using 10 channels on different microfluidic devices, with variations of 1.4 % in depth and 1.8% in width. Fabrication of microchannels on polymethylmethacrylate (PMMA) substrates using novel microfabrication methods is demonstrated. The image of microchannels is transferred from a silicon master possessing the inverse image of the microchannel to a PMMA plate by using hot embossing methods. The silicon master is electrostatically bonded to a Pyrex 7740 glass wafer, which improves the device yield from about 20 devices to hundreds of devices per master. Effects of embossing temperature, pressure and time on the accuracy of replication are systematically studied using the orthogonal factorial design. According to the suggested experimental model , the time for the whole embossing procedure is shorten from about 20 min to 6 min and the accuracy of replication is 99.3%. The reproducibility of the hot embossing method is evaluated using 10 channels on different microfluidic devices, with variations of 1.4% in depth and 1.8% in width.