Ultrasound (US) imaging has become an essential modality in preclinical research due to its numerous advantages in the study of biological systems and disease models. This non-invasive, real-time imaging technique enables high-resolution, dynamic visualization of soft tissues, vasculature, and organ structures, positioning it as a critical radiological tool in the fields of oncology and cardiology. This book chapter reviews the application of various US imaging techniques in preclinical research. Specifically, it examines anatomical imaging modes, including 2D and 3D B-mode US, for assessing disease progression, evaluating therapeutic interventions, monitoring the effects of novel drugs or treatments, and guiding image-assisted injections. The chapter further explores contrast-enhanced imaging techniques for evaluating tissue perfusion. Additionally, M-mode, Strain and Strain Rate imaging, 4D echocardiography, and Doppler modalities are discussed in the context of assessing heart chamber dimensions, contractility, blood flow velocities, and valve function, contributing to a comprehensive evaluation of cardiac health. Collectively, US imaging serves as a versatile, cost-effective, and reproducible platform, enhancing our understanding of disease mechanisms, supporting drug development, and advancing translational research.