Acoustic particle manipulation in microfluidic channels is becoming a powerful tool in microfluidics to control micrometer sized objects in medical, chemical and biological applications. By creating a standing acoustic wave in the channel, the resulting pressure field can be employed to trap or sort particles. To design efficient and reproducible devices, it is important to characterize the pressure field throughout the volume of the microfluidic device. Here, we used an optically trapped particle as probe to measure the forces in all three dimensions. By moving the probe through the volume of the channel, we imaged spatial variations in the pressure field. In the direction of the standing wave this revealed a periodic energy landscape for 2 μm beads, resulting in an effective stiffness of 2.6 nN m(-1) for the acoustic trap. We found that multiple fabricated devices showed consistent pressure fields. Surprisingly, forces perpendicular to the direction of the standing wave reached values of up to 20% of the main-axis-values. To separate the direct acoustic force from secondary effects, we performed experiments with different bead sizes, which attributed some of the perpendicular forces to acoustic streaming. This method to image acoustically generated forces in 3D can be used to either minimize perpendicular forces or to employ them for specific applications in novel acoustofluidic designs.