The introduction of optoelectrokinetics (OEK) into lab-on-a-chip systems has facilitated a new cutting-edge technique-the OEK-based micro/nanoscale manipulation, separation, and assembly processes-for the microfluidics community. This technique offers a variety of extraordinary advantages such as programmability, flexibility, high biocompatibility, low-cost mass production, ultralow optical power requirement, reconfigurability, rapidness, and ease of integration with other microfluidic units. This paper reviews the physical mechanisms that govern the manipulation of micro/nano-objects in microfluidic environments as well as applications related to OEK-based micro/nanoscale manipulation-applications that span from single-cell manipulation to single-molecular behavior determination. This paper wraps up with a discussion of the current challenges and future prospects for the OEK-based microfluidics technique. The conclusion is that this technique will allow more opportunities for biomedical and bioengineering researchers to improve lab-on-a-chip technologies and will have far-reaching implications for biorelated researches and applications in the future.