Abstract
PURPOSE: This study aimed to develop a high-resolution full-field optical coherence tomography (HR-FFOCT) system for automated three-dimensional (3D) imaging of cultivated epithelial cell sheets and biopsied tissue used in corneal regenerative therapy. MATERIALS AND METHODS: A commercial HR-FFOCT system (ApolloVue® S100, AMO, Taiwan), originally for dermatological imaging, was re-engineered for <2 μm resolution imaging of rabbit limbal and oral mucosal tissues for limbal stem cell deficiency treatment. Modifications included a piezoelectric transducer for precise Z-stack acquisition, customized LED illumination for registration, and a specialized platform for culture dishes. The system enabled en face and cross-sectional imaging with 3D reconstruction. Rabbit-derived products for cultivated limbal epithelial transplantation (CLET) and cultivated oral mucosal epithelial transplantation (COMET) were imaged before cultivation. Morphology, stratification, and protein expression were analyzed and validated with immunocytochemistry (ICC). RESULTS: The optimized system produced high-resolution en face and Z-stack images with accurate alignment, capturing stratified epithelial layers in CLET and oral mucosal tissue. Reconstructed 3D images revealed structural detail consistent with ICC-verified expression of junctional proteins, including occludin and actin. Both two-dimensional and 3D visualization of biopsied oral mucosal tissue was achieved. The system enabled noninvasive monitoring of epithelial sheet architecture and thickness without sectioning. CONCLUSION: The reconstructed HR-FFOCT system provides a noninvasive, real-time imaging platform for assessing epithelial cell sheets and biopsied tissue in corneal regenerative therapy. It offers potential for standardizing quality evaluation of cell-based products before transplantation and advancing translational applications in regenerative medicine.