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Optical fiber tweezers offer a simple, low-cost and portable solution for non-invasive trapping and manipulation of particles. However, single-fiber tweezers require fiber tip modification (tapering, lensing, etc.) and the dual-fiber approach demands strict alignment and positioning of fibers for robust trapping of particles. In addition, both tweezing techniques offer a limited range of particle manipulation and operate in low flow velocity regimes (a few 100 μm s−1) when integrated with microfluidic devices. In this paper, we report a novel opto-hydrodynamic fiber tweezers (OHT) platform that exploits the balance between the hydrodynamic drag force and optical scattering forces to trap and manipulate single or multiple particles of various shapes, sizes, and material compositions in a microfluidic channel. 3D hydrodynamic flow focusing offers an easy and dynamic alignment of the particle trajectories with the optical axis of the fiber, which enables robust trapping of particles with high efficiency of >70% and throughput of 14 particles per minute (operating flow velocity: 1000 μm s−1) without the need for precision stages or complex fabrication. By regulating the optical power and flow rates, we were able to trap single particles at desired positions in the channel with a precision of ±10 μm as well as manipulate them over a long range upstream or downstream with a maximum distance of 500 μm. Our opto-hydrodynamic tweezers offer an alternative to conventional optical fiber tweezers for several applications in physics, biology, medicine, etc. [ABSTRACT FROM AUTHOR]