Intravital microscopy (IVM) and optical coherency tomography (OCT) are powerful optical imaging tools that allow visualization of dynamic biological activities in living subjects with subcellular resolutions. They have been used in preclinical and clinical cancer imaging, providing insights into the complex physiological, cellular, and molecular behaviors of tumors. They have revolutionized cancer diagnosis and therapies, allowing for real-time observation of biologic processes in vivo, including angiogenesis and immune cell interactions. Recent developments in techniques for observing deep tissues of living animals have improved bioluminescent proteins, fluorescent proteins, fluorescent dyes, and detection technologies like two-photon excitation microscopy. These technologies have become indispensable tools in basic sciences, preclinical research, and modern drug development. In Vivo imaging can detect subcellular signaling or metabolic events in living animals, but depth-dependent signal attenuation limits the depth from which significant data can be obtained. Cancer cell motility and invasion are key features of metastatic tumors, but only a small portion of tumor cells are motile and metastasize due to genetic, epigenetic, and microenvironmental heterogeneities.