In recent years, dielectrophoretic force has been used to manipulate colloids, inert particles, and biological microparticles, such as red blood cells, white blood cells, platelets, cancer cells, bacteria, yeast, microorganisms, proteins, DNA, etc. This specific electrokinetic technique has been used for trapping, sorting, focusing, filtration, patterning, assembly, and separating biological entities/particles suspended in a buffer medium. Dielectrophoretic forces acting on particles depend on various parameters, for example, charge of the particle, geometry of the device, dielectric constant of the medium and particle, and physiology of the particle. Therefore, to design an effective micro-/nanofluidic separation platform, it is necessary to understand the role of the aforementioned parameters on particle motion. In this paper, we review studies particularly related to dielectrophoretic separation in microfluidic devices. Both experimental and theoretical works by several researchers are highlighted in this article covering AC and DC DEP. In addition, AC/DC DEP, which uses a combination of low frequency AC and DC voltage to manipulate bioparticles, has been discussed briefly. Contactless DEP, a variation of DC DEP in which electrodes do not come in contact with particles, has also been reviewed. Moreover, dielectrophoretic force-based field flow fractionations are featured to demonstrate the bioparticle separation in microfluidic device. In numerical front, a comprehensive review is provided starting from the most simplified effective moment Stokes-drag (EMSD) method to the most advanced interface resolved method. Unlike EMSD method, recently developed advanced numerical methods consider the size and shape of the particle in the electric and flow field calculations, and these methods provide much more accurate results than the EMSD method for microparticles.