The development of lab-on-a-chip (LOC) devices over the past decade has attracted growing interest. LOC devices aim to achieve the miniaturization, integration, automation and parallelization of biological and chemical assays. One of the applications, the ability to effectively and accurately manipulate and separate micro- and nano-scale particles in an aqueous solution, is particularly appealing in biological, chemical and medical fields. Among the technologies that have been developed and implemented in microfluidic microsystems for particle manipulation and separation (such as mechanical, inertial, hydrodynamic, acoustic, optical, magnetic and electrical methodologies), dielectrophoresis (DEP) may prove to be the most popular because of its label-free nature, ability to manipulate neutral bioparticles, analyse with high selectivity and sensitivity, compatibility with LOC devices, and easy and direct interface with electronics. The required spatial electric non-uniformities for the DEP effect can be generated by patterning microelectrode arrays within microchannels, or placing insulating obstacles within a microchannel and curving the microchannels. A wide variety of electrode- and insulator-based DEP microdevices have been developed, fabricated, and successfully employed to manipulate and separate bioparticles (i.e. DNA, proteins, bacteria, viruses, mammalian and yeast cells). This review provides an overview of the state-of-the-art of microfabrication techniques and of the structures of dielectrophoretic microdevices aimed towards different applications. The techniques used for particle manipulation and separation based on microfluidics are provided in this paper. In addition, we also present the theoretical background of DEP.