Single-cell rotation is a fundamental manipulation used in a wide range of biotechnological applications such as cell injection and enucleation. However, there are currently few methods for the 3D rotation of single cells. Here, this chapter presents different biochip platforms based on a dielectrophoresis technique to achieve 3D rotation. In-plane (yaw) and out-of-plane rotation (pitch) can be achieved by applying different AC signal configurations, respectively. This use of 3D rotation facilitates several applications. For example, in-plane rotation can be used to measure the rotation spectra, and this can be used to estimate the dielectric parameters. The out-of-plane rotation can help reconstruct 3D cell models.