During particle (e.g., a biological cell)–laser interaction, optical force is generated from the universal momentum exchange; the extent of the exchange reflects the intrinsic properties of the particle. Consequently, optical force has been proposed for use in particle separation: There are several optical force-based, label-free methods, each of which is based on the fact that a change in shape, composition, internal structure, or size causes a change in a particle’s optical properties, and thus the optical force experienced by the particle. Although in most methods, the measured parameters are not sensitive to cell-type changes and the accompanying changes in optical forces: we previously10 demonstrated that the laser-guidance technique, which measures the speed of the optical force-driven cell motion, is highly sensitive in cell detection. For example, modifying a single gene in a mouse lung-carcinoma tumor cell can cause approximately a 40% change in guidance speed. In this study, we applied laser guidance in a microfluidic biochip that provided the possibility of high-throughput cell separation after laser guidance-based cell detection.