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Opt. Express"],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:p>Structural and functional assessment of human cardiac organoids (hCOs) is essential for pharmaceutical drug screening and disease modeling. Existing evaluation approaches have provided valuable insights, but many rely on invasive sensors that may influence contractility, or on 2D imaging approaches that are constrained by geometric assumptions and restrict field of view. To address these challenges, we present a non-invasive imaging approach using 4D optical coherence tomography (4D-OCT) to reconstruct a synchronized, volumetric video of beating hCOs by denoising, segmenting, and synchronizing adjacent time-series B-scan stacks. We used 4D-OCT to visualize the morphological deformation of beating hCOs, enabling quantification of chamber deformation ratio (CDR) and deformation fraction (DF). Ultimately, our 4D-OCT pipeline measured an 8-fold higher DF in the vascular endothelial growth factor-treated hCO compared to the control hCO for proof of concept. 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