Ultrasound localization microscopy (ULM) has superior spatial resolution; however, it requires contrast agents and long data acquisition and processing time. Null subtraction imaging (NSI) is a nonlinear beamforming technique that improves the spatial resolution and reduces grating lobes with low computational cost. We combined pulse-inversion (PI) nonlinear imaging with NSI to increase resolution. Pulses with center frequency of 10.42 MHz and its inverted version were transmitted to obtain the second harmonic at 20.84 MHz (wavelength of ∼74 μm). The transducer pitch was 35% larger than the wavelength of the receive signal, which would introduce grating lobes in the field of view. A total of 1000, 9-angle (−8 to 8 degree in 2 degrees step) coherently compounded frames were acquired in 1 s. SVD filters were applied in raw RF data to filter out tissue signal. The dc offset was set to 0.1 in NSI imaging. Grating lobes, which were obvious in DAS images, were removed in the NSI images. Higher spatial resolution and contrast were observed from the NSI microvessel images. The spatial resolution of the NSI images using harmonic imaging approached one fourth of a wavelength with computation time increased by only 40% compared to DAS.
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