Abstract:We present phase subtraction imaging (PSI), a new spatial-temporal beamforming method that enables micrometer level resolution imaging of microvessels in live animals without labels, which are microbubbles in ultrasound super-resolution imaging. Subtraction of relative phase differences between consecutive frames beamformed with mismatched apodizations is used in PSI to overcome the diffraction limit. We validated this method by imaging both the mouse brain and rabbit kidney using different ultrasound probes and scanning machines.