Measurement of blood flow velocity is key to understanding physiology and pathology in vivo. While most measurements are performed at the middle of the blood vessel, little research has been done on characterizing the instantaneous blood flow velocity distribution. This is mainly due to the lack of measurement technology with high spatial and temporal resolution. Here, we tackle this problem with our recently developed dual-wavelength line-scan third-harmonic generation (THG) imaging technology. Simultaneous acquisition of dual-wavelength THG line-scanning signals enables measurement of blood flow velocities at two radially symmetric positions in both venules and arterioles in mouse brain in vivo. Our results clearly show that the instantaneous blood flow velocity is not symmetric under general conditions.