The manipulation of particles in fluids using microfluidic devices is a fundamental task in Lab-on-a-Chip applications. Grooved structures have been widely studied in particle handling and fluid mixing in microfluidic channel systems. In this study, we report use of patterning flows produced by a series of grooved surfaces with different geometrical setups integrated into a microfluidic device, to continuously manipulate the flowing particles, ranging from 6 to 20 μm in diameters, of comparable sizes to the depth of the channel. COMSOL, a multiphysics modeling software that can help predict engineering trends, is used to systematically quantify the following parameters: 1) channel depth, 2) groove width, 3) groove depth, 4) groove angle, and 5) flow speed, which may affect the performance of separation for flowing particles inside the channel. The device is fabricated using softlithographic techniques and is composed of inlets, microfluidic channels, and outlets for loading, manipulating and retrieving cell suspensions, respectively. Experimental results indicated that the particles were evenly distributed in the entrance of the microchannel and illustrate patterns of enriching, focusing, or size-selective profiles after passing through the grooved area. The preliminary simulation results also demonstrated that particles tend to bias towards the sidewall after flowing through the grooves.