The magnitude and direction of the ultrasonic radiation forces that act on individual particles in a standing-wave field were determined using a microscope-based imaging system. The forces are calculated from measured particle velocities assuming that the drag force, given by Stokes’ law, is exactly counterbalanced by the imposed ultrasonic forces. The axial primary radiation force was found to vary sinusoidally with axial position and to be proportional to the local acoustic energy density, as predicted by theory. The magnitude of the transverse primary force was determined by two independent methods to be about 100-fold weaker than the axial force. Separation concepts exploiting the transverse force for cell retention have been successful despite the great disparity in magnitude between the axial and transverse-force components. This may be explained by the reduced hydrodynamic forces on aggregated particles in transverse flow due to their alignment in the sound field.