This paper presents a highly efficient single cell capture scheme using hydrodynamic guiding structures in a microwell array. The implemented structure has a capturing efficiency of >80%, and has a capacity to place individual cells into separated microwells, allowing for the time-lapse monitoring on single cell behavior. Feasibility was tested by injecting microbeads (15 μm in diameter) and prostate cancer PC3 cells in an 8×8 microwell array chip and >80% of the microwells were occupied by single ones. Using the chips, the number of cells required for cell assays can be dramatically reduced and this will facilitate overcoming a huddle of assays with scarce supply of cells.