This paper demonstrates the utilization of 3D semispherical shaped microelectrodes for dielectrophoretic manipulation of yeast cells. The semispherical microelectrodes are capable of producing strong electric field gradients, and in turn dielectrophoretic forces across a large area of channel cross-section. The semispherical shape of microelectrodes avoids the formation of undesired sharp electric fields along the structure and also minimizes the disturbance of the streamlines of nearby passing fluid. The advantage of semispherical microelectrodes over the planar microelectrodes is demonstrated in a series of numerical simulations and proof-of-concept experiments aimed toward immobilization of viable yeast cells.