Light sheet fluorescence microscopy (LSFM) has emerged as a transformative imaging technique in the study of drug delivery and embryonic development, offering high-resolution, real-time visualization with minimal phototoxicity. This review examines the application of LSFM in tracking drug pharmacokinetics, tissue-specific targeting, and drug efficacy during critical phases of embryonic development. Recent advancements in fluorescent labeling and machine learning integration have enabled more precise monitoring of drug release, distribution, and interaction with developing tissues. The ability of LSFM to capture long-term dynamics at single-cell resolution has revolutionized drug discovery, especially in nanomedicine and targeted therapies. By integrating LSFM with multimodal imaging and AI-driven data analysis, researchers are now better equipped to explore complex biological processes and optimize drug delivery in a highly controlled, minimally invasive manner. Finally, the review highlights the pivotal role of LSFM in advancing drug delivery research, addressing existing challenges, and unlocking new frontiers in clinical applications.