Microbubbles are used as effective contrast agents in ultrasound imaging. However, low stability of the microbubbles limits their use for prolonged period of time in medical applications. The aim of this dissertation is to increase the stability of microbubbles under ultrasound. The stability and acoustic response of microbubbles were investigated under ultrasound as a function of their shell composition. Microbubbles were fabricated using combinations of phospholipid (DSPC) and an emulsifier (PEG40St) in different molar ratios. It was found that adding the emulsifier decreased the microbubble stability under ultrasound; however, the echogenicity of microbubbles was shown to increase with increasing emulsifier content. A method was developed to estimate the concentration of microbubbles with ultrasound. Hydrostatic pressure studies showed that the microbubbles recovered their spherical structures at low pressure pulses, in contrast, disappeared in a very short time at high pressure pulses. B-mode ultrasound intensity of microbubbles was investigated at different ultrasound powers under Doppler ultrasonography, and for the first time, a model was developed to relate the intensity to effective bubble concentration. We calculated acoustic energy thresholds and explained a possible mechanism for the destruction of microbubbles under ultrasound. The effect of shell loadings on the acoustic response and stability of microbubbles were investigated under ultrasound. It was found that both the echogenicity and stability of microbubbles increased with increasing mass of the loadings on microbubble shell. In-vivo studies showed that the acoustic performance of in-house made microbubbles was comparable to that of commercial standard Vevo MicroMarker® contrast agents.