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The change in direction of a cell flowing over an oblique micro groove has been analyzed in vitro. The micro flow-channel (0.05 mm height × 1 mm width × 25 mm length) with oblique micro grooves (4.5 μm depth) was manufactured on a polydimethylsiloxane (PDMS) disk by the micromachining technique. The angle between the main flow direction and the longitudinal axis of the groove is 45 degrees. The effect of variation of the groove width (0.03 mm, 0.04 mm, and 0.05 mm) was studied. Myoblasts (C2C12: mouse myoblast cell line) were used in the test. The main flow velocity (0.02 mm/s < vx < 0.23 mm/s) of the medium was controlled by the pressure difference between the inlet and the outlet. The shape of each flowing cell was tracked on a movie recorded by the camera attached to the eyepiece of the microscope. The experimental results show that the change of the direction of each cell by each groove depends on the shape of the cell, which depends on both the shape of the cell and the width of the groove.