The aim of this paper is to develop a novel method to separate Microthrix parvicella (M. parvicella) filaments from activated sludge easily and quickly, as there are a few difficulties in the isolation of M. parvicella filaments, such as complicated isolation process, time consuming, etc. In this work, a series of hydrophobic plate with and without microchannels have been prepared for the separation of M. parvicella filaments. The results showed that the presence of microchannels and hydrophobic property of the hydrophobic plates affected the separation efficiency of M. parvicella significantly. The scanning electron microscope and Keyence Digital Microscope analysis results showed that the diameter of microchannels was similar to the width of M. parvicella filament, which was beneficial for the fastening of M. parvicella filaments on the plate. The hydrophobic property of the prepared plates was tested by contact angle of water droplets, and the results displayed that the polydimethylsiloxane (PDMS) plate possessed the highest contact angle compared with that of other plates, like polymethylmethacrylate, polystyrene plate, and PDMS plate with no hydrophobic microchannels. Thus, it was concluded that the high separation efficiency of PDMS plates to M. parvicella filaments was due to its best hydrophobic property.