Grooved structures have been widely studied in particle separation and fluid mixing in microfluidic channel systems. In this brief report, we demonstrate the use of patterning flows produced by two different sorts of grooved surfaces: single slanted groove series (for enrichment patterns) and V-shaped groove series (for focusing patterns), into a microfluidic device to continuously manipulate the flowing particles, including microbeads with , , and in diameter and mouse dendritic cells of comparable sizes to the depth of the channel. The device with grooved channels was developed and fabricated by soft-lithographic techniques. The particle distributions after passing through the single slanted grooves illustrate the size-dependent enrichment profiles. On the other hand, particles passing through the V-shaped grooves show focusing patterns downstream, for the combination effect from both sides of single slanted grooves setup side-by-side. Compared with devices utilizing sheath flows, the focusing patterns generated in this report are unique without introducing additional flow control. The alignment of the concentrated particles is expected to facilitate the visualization of sizing and counting in cell-based devices. On the other hand, the size-dependent patterns of particle distributions have the potential for the application of size-based separation.