Nonlinear Frequency Mixing Ultrasound Imaging of Nanoscale Contrast Agents.

OBJECTIVE: Nanoscale ultrasound contrast agents show promise as alternatives for diagnostics and therapies due to their enhanced stability and ability to traverse blood vessels. Nonetheless, their reduced size limits echogenicity. This study introduces an enhanced nanobubble frequency mixing ultrasound imaging method, by capitalizing on their nonlinear acoustic response to dual-frequency excitation. METHODS: A single broadband transducer (L12-3v) controlled by a programmable ultrasound system was used to transmit a dual-frequency single-cycle wavefront. The frequency mixing effect enabled simultaneous transducer capture of nanobubble-generated sum and difference frequencies in real time without the need for additional hardware or post-processing, by substituting the single-frequency wavefront in a standard contrast harmonic pulse inversion imaging protocol, with the dual-frequency wavefront. RESULTS: Optimization experiments were conducted in tissue mimicking phantoms. Among the dual-frequency combinations that were tested, the highest contrast was obtained using 4&8 MHz. The nanobubble contrast improved with increased mechanical index, and achieved a maximal contrast improvement of 8.4 ± 0.5 dB compared to 4 MHz pulse inversion imaging. In imaging of a breast cancer tumor mouse model, after a systemic nanobubble injection, the contrast was improved by 3.4 ± 1.7, 4.8 ± 1.8, and 6.3 ± 1.6 dB for mechanical indices of 0.04, 0.08, and 0.1, respectively. CONCLUSION: Nonlinear frequency mixing significantly improved the nanobubble contrast, which facilitated their imaging in-vivo. SIGNIFICANCE: This study offers a new avenue to enhance ultrasound imaging utilizing nanobubbles, potentially leading to advancements in other diagnostic applications.

Ultrasound Frequency Mixing for Enhanced Contrast Harmonic Imaging of Microbubbles.

Microbubbles (MBs) serve as contrast agents in diagnostic ultrasound (US) imaging. Contrast harmonic imaging (CHI) of MBs takes advantage of their nonlinear properties that generate additional harmonic frequencies in the received spectrum. However, CHI suffers from limitations in terms of contrast, the signal-to-noise ratio, and artifacts. This article presents an enhanced, real-time, nonlinear imaging technique based on the excitation of MBs with a dual frequency waveform. The MBs trigger a frequency mixing effect that generates additional frequency components in the received spectrum; i.e., difference and sum frequencies, in addition to the standard harmonics, thus amplifying the MB's nonlinear response and enhancing image contrast. In this real-time approach, two single frequency waveforms are superpositioned into a dual frequency transmission. The dual frequency waveform is incorporated into a standard pulse-inversion (PI) sequence and is transmitted by an array transducer using an arbitrary waveform generator (AWG) in a programmable US system. Upon receive, standard dynamic receive beamforming is used, without additional post processing. Numerical simulations using the Marmottant model are used to confirm the generation of the difference frequency in the MB's backscattered echoes. The resulting image quality enhancement is demonstrated in a tissue-mimicking phantom containing MBs' suspension. A maximal contrast improvement of 3.43 dB compared to standard PI was achieved, along with a reduction by 4.5 fold in the mechanical index (MI).