In present study, the behavior of electrospun ZnO nanofibers under the application of low frequency alternating (AC) electric fields within the range of 1 Hz-20 kHz was investigated using planar parallel electrodes. In the first stage, ZnO nanofibers with diameters up to 100 nm were prepared by electrospinning. At 1 Hz, the strong electric field-induced fluid flow due to charge redistribution around the electrode swept the fibers on the electrode surface farther from the edges. As the frequency was increased to 1 kHz, most depositing fibers accumulated on the electrode edges and in the meantime, a small fraction of them was pushed into the surface as a result of AC electroosmosis effect. Above 1 kHz, the fibers were assembled within the electrode gap bridging the interelectrode space. The dielectrophoresis force was considered responsible for the assembly and relative alignment of ZnO nanofibers above 1 kHz.