The harmonic generation due to the nonlinear behavior of Ultrasound Contrast Agents (UCAs) must be exploited for improved efficiency when imaging vascular targets in the neighborhood of highly echogenic tissue. One may even further improve the efficiency by focusing on the subharmonic generation of the UCAs, which is an even more exclusive property than the generation of higher harmonics, for improved Contrast-to-Tissue ratio (CTR). The aim of this work was first, the design of a set-up for nonlinear imaging of Poly-Vinyl Alcohol (PVA) based UCAs on The Verasonics Research System with special focus on nondestructive Subharmonic Imaging. The second part of the work addressed the evaluation of the subharmonic response provided by the agents in the developed setup. Six different imaging techniques were developed. These were Fundamental B-mode imaging (FB), Pulse Inversion imaging (PI), and a Contrast Pulse Sequence based on three pulses (CPS3), with and without a focus on the subharmonic component by the implementation of a Linear Bandpass Filter (LBF). Experiments were performed on a tissue mimicking flow phantom and the performance of the agents for each technique was determined in terms of CTR and CNR. The PVA agents provided a backscattering enhancement of the order of 23 dB through FB imaging. However, the performance of the FB technique was unsatisfactory in terms of CTR. The CPS3 sequence performed best of the six techniques with an improvement of 14 dB and 13 dB in CTR and CNR, respectively, compared with the FB technique. Combining the LBF around the subharmonic component with the multi-pulse techniques of PI and CPS3 resulted in a degraded CTR performance due to significant amount of signals from tissue around the subharmonic component and insufficient subharmonic detection from the PVA agents.

Evolution of vulnerable carotid plaques are crucial reason for cerebral ischemic strokes and identifying them in the early stage can become very important in avoiding the risk of stroke. In order to improve the identification and quantification accuracy of infancy plaques better visualization techniques are needed. Improving the visualization and quantification of neovascularization in carotid plaque using contrast enhanced ultrasound imaging still remains a challenging task. In this thesis work, three optimization techniques are proposed, which showed an improvement in the sensitivity of contrast agents when compared to the conventional clinical settings and insonation strategies. They are as follows:1) Insonation at harmonic specific (2nd harmonic) resonance frequency instead of resonance frequency based on maximum energy absorption provides enhanced nonlinear contribution.2) At high frequency ultrasound imaging, shorter pulse length will provide improved harmonic signal content when compared to longer pulse lengths. Applying this concept to multi- pulse sequencing (Pulse Inversion and Cadence contrast pulse sequencing) resulted in increased magnitude of the remaining harmonic signal after pulse summations.3) Peak negative pressure optimization of Pulse Inversion and Cadence contrast pulse sequencing was showed to further enhance the nonlinear content of the backscattered signal from contrast microbubbles without increasing the safety limits, defined by the mechanical index.The results presented in this thesis are based on computational modeling (Bubblesim software) and as a future continuation we plan to verify the simulation results with vitro studies.