This paper introduces microfluidic manipulation of microorganism by opto-electrokinetic technique, named rapid electrokinetic patterning (REP). REP is a hybrid method that utilizes the simultaneous application of a uniform electric field and a focused laser to manipulate various kinds and types of colloidal particles. Using the technique in preliminary experiments, we have successfully aggregated, translated, and trapped not only spherical polystyrene, latex, and magnetic particles but also ellipsoidal glass particles. Extending the manipulation target to cells, we attempted to manipulate saccharomyces cerevisiae (S. cerevisiae), the most commonly used microorganism for food fermentation and biomass production. As a result, S. cerevisiae were assembled and dynamically trapped by REP at arbitrary location on an electrode surface. It firmly establishes the usefulness of REP technique for development of a high-performance on-chip bioassay system.