Advanced imaging strategies in cardiac organoids: bridging the gap between structural complexity and functional analysis

Cardiac organoids have emerged as pivotal models for cardiovascular disease research and drug development due to their ability to recapitulate the in vivo cardiac microenvironment, structure, and function. Imaging technologies are core tools for deciphering their three-dimensional (3D) complexity. This review outlines the key framework of cardiac organoid imaging research, focusing on pre-imaging sample processing, mainstream imaging technologies, image analysis workflows and applications in disease mechanisms and drug screening. We also provide guidelines for technique selection based on research objectives. Currently, long-term 4D imaging of cardiac organoids is still in its infancy. Future efforts should optimize imaging strategies and advance high-resolution dynamic techniques to deepen understanding of the temporal dynamics of cardiac pathology, providing technical support for cardiovascular research.