Investigating shear flows is important in technical applications as well as in fundamental research. Velocity measurements with high spatial resolution are necessary. Laser Doppler anemometry allows nonintrusive precise measurements, but the spatial resolution is limited by the size of the measurement volume to approximately 50 microm. A new laser Doppler profile sensor is proposed, enabling determination of the velocity profile inside the measurement volume. Two fringe systems with contrary fringe spacing gradients are generated to determine the position as well as the velocity of passing tracer particles. Physically discriminating between the two measuring channels is done by a frequency-division-multiplexing technique with acousto-optic modulators. A frequency-doubled Nd:YAG laser and a fiber-optic measuring head were employed, resulting in a portable and flexible sensor. In the center of the measurement volume of approximately 1-mm length, a spatial resolution of approximately 5 microm was obtained. Spatially resolved measurements of the Blasius velocity profile are presented. Small velocities as low as 3 cm/s are measured. The sensor is applied in a wind tunnel to determine the wall shear stress of a boundary layer flow. All measurement results show good agreement with the theoretical prediction.