目的：开发细胞级高分辨率、结构与功能一体化的双模态全视场光学相干层析系统(full-field optical coherence tomography,FFOCT)，实现角膜缘组织的双模态FFOCT成像。方法：基于Linnik干涉成像原理，利用高数值孔径显微物镜(NA=0.8)及高速平面互补金属氧化物半导体(complementary metal oxide semiconductor,CMOS)相机，设计搭建高分辨率的组织静态结构和内源动态功能成像一体化双模态FFOCT系统；构建基于四相位调制结构影像提取及时域干涉信号动态频谱分析的功能影像重建算法；对人供体角膜缘组织开展各深度层的双模式FFOCT成像有效性验证。结果：搭建的双模态FFOCT成像系统可实现横向分辨率0.5 μ m，轴向分辨率1.7 μ m，成像视野320 μ m×320 μ m，相机采集速度100 Hz。系统实现角膜缘组织无外源标记情况下的细胞级分辨率三维结构和内源功能成像，FFOCT静态结构影像清晰显示角膜缘上皮、Vogt栅栏、隐窝、基质、血管及淋巴管等结构，FFOCT动态功能影像突出显示了代谢活跃细胞(角膜缘上皮细胞、免疫细胞等)。结论：双模态FFOCT高分辨率成像系统可提供角膜缘微观结构和活细胞无标记内源功能可视化信息，将为角膜缘疾病的研究及临床诊疗提供全新的成像分析技术。
Objective: To develop a cellular-level, high-resolution, integrated dual-modal full-field optical coherence tomography (FFOCT) system capable of simultaneously imaging the structure and function of limbus tissue. Methods: Utilizing the Linnik interference imaging principle, a high-resolution dual-modal FFOCT system was designed and constructed using a high numerical aperture (NA=0.8) microscope objective and a high-speed flat CMOS camera. A functional imaging reconstruction algorithm based on four-phase modulation structure image extraction and dynamic frequency spectrum analysis of temporal interference signals was developed. The effectiveness of dual-mode FFOCT imaging at various depth layers of human corneal limbal tissue was validated. Results: The constructed dual-modal FFOCT imaging system achieved lateral resolution of 0.5 μ m, axial resolution of 1.7 μ m, imaging field of view of 320 μ m × 320 μ m, and camera acquisition speed of 100 Hz. The system enabled cellular-level resolution three-dimensional structural and intrinsic functional imaging of corneal limbal tissue without exogenous labeling. Static structural FFOCT images clearly displayed limbal epithelium, palisades of Vogt, crypts, stroma, blood vessels, and lymphatic vessels, while dynamic functional FFOCT images highlighted metabolically active cells (limbal epithelial cells, immune cells, etc.). Conclusion: The dual-modal FFOCT high-resolution imaging system provides visualization of corneal limbal microstructural and live cell intrinsic functional information without labeling, offering a novel imaging analysis technique for research and clinical diagnosis and treatment of limbal diseases.
角膜缘的细胞，特别是角膜缘干细胞，对于维持角膜的透明和健康至关重要。基于影像技术对角膜缘进行高精度可视化评价是相关疾病诊疗的重要手段。眼科临床使用的裂隙灯显微镜、共聚焦显微镜、眼前节光学相干断层扫描仪(optical coherence tomography,OCT)等成像技术，因低分辨、低对比度、侵入性等原因，限制了其在角膜缘细胞结构及功能影像评估中的应用。本团队创新研发新型双模态全视场光学相干断层扫描仪(full-field OCT，FFOCT)，成功实现了无标记的角膜缘细胞级分辨率结构及功能成像。FFOCT基于空间非相干光平面干涉原理提取组织内部散射光，获得微米级分辨率三维结构成像；通过FFOCT原始相干信号的高时空分辨率采集及动态特征解析，实现源于活细胞新陈代谢运动的无标记细胞功能影像可视化。双模态FFOCT创新性地整合了高分辨率、无标记的结构及功能成像模态，不仅清晰获取角膜缘组织的高精结构特征如Vogt栅栏、角膜缘隐窝、血管壁等，同时还能捕捉不同角膜缘细胞内的代谢活性动态变化，无需使用外源荧光染料或标记剂，为角膜缘生物学及疾病机制研究提供全新细胞水平结构及功能成像方法，具有广泛应用前景。
角膜缘的细胞，特别是角膜缘干细胞，对于维持角膜的透明和健康至关重要。基于影像技术对角膜缘进行高精度可视化评价是相关疾病诊疗的重要手段。眼科临床使用的裂隙灯显微镜、共聚焦显微镜、眼前节光学相干断层扫描仪(optical coherence tomography,OCT)等成像技术，因低分辨、低对比度、侵入性等原因，限制了其在角膜缘细胞结构及功能影像评估中的应用。本团队创新研发新型双模态全视场光学相干断层扫描仪(full-field OCT，FFOCT)，成功实现了无标记的角膜缘细胞级分辨率结构及功能成像。FFOCT基于空间非相干光平面干涉原理提取组织内部散射光，获得微米级分辨率三维结构成像；通过FFOCT原始相干信号的高时空分辨率采集及动态特征解析，实现源于活细胞新陈代谢运动的无标记细胞功能影像可视化。双模态FFOCT创新性地整合了高分辨率、无标记的结构及功能成像模态，不仅清晰获取角膜缘组织的高精结构特征如Vogt栅栏、角膜缘隐窝、血管壁等，同时还能捕捉不同角膜缘细胞内的代谢活性动态变化，无需使用外源荧光染料或标记剂，为角膜缘生物学及疾病机制研究提供全新细胞水平结构及功能成像方法，具有广泛应用前景。
目的：分析人眼的睑板腺形态学特征，探索睑板腺分析系统在眼表疾病的应用研究。方法：中山眼科中心入组正常受试者24例(42眼)，进行睑板腺红外摄影。选取受试者中的10例(20眼)在同型号的设备上由二名操作员分别进行睑板腺红外摄影。图像通过自行设计的分析软件对上睑结膜中央5条腺体形态学参数进行定量分析，对数据进行重复性测试。结果：测量的生物参数腺体直径为(0.48±0.09) mm，腺体长度为(5.25±0.68) mm，腺体面积为(2.12±0.53) mm，腺体形变系数为10.01±3.85，显影值为6.32±1.23，中央五条腺体占中央区域面积百分比为(10.94±2.20)%，腺体占上睑结膜面积百分比为(58.07±8.13)%。各指标两次测量值差异无统计学意义(P>0.05)。重复性分析结果显示：腺体各项生物参数的变异系数(coefficients of variation，CV)均小于5%，组内变异系数(intraclass correlation coefficient，ICC)均大于0.95。结论：睑板腺综合分析系统对腺体的形态学分析有良好的可靠性和一致性，有望为临床上对睑板腺腺体功能评估提供新的非侵入性参考指标。
Objective: To analyze the morphological characteristics of meibomian glands in human eyes and to explore the application research of meibomian glands analysis system in ocular surface diseases. Methods: A total of 24 healthy subjects were recruited by Zhongshan Ophthalmic Center to infrared photography of meibomian glands. Ten of healthy subjects were selected by the two operators for infrared photography of meibomian glands on the same model of equipment. The images were repeatedly measured and analyzed by the self-designed analysis software on the morphological measurements of the five glands in the center of the upper eyelid. Results: The measured biological parameters are shown below: the average gland diameter was (0.48±0.09) mm, the average gland length was (5.25±0.68) mm, the average gland area was (2.12±0.53) mm, the gland deformation coefficient was 10.01±3.85, the development value was 6.32±1.23, the percentage of the five central glands in the central area was (10.94±2.20)%, and the glands accounted for (58.07±8.13)% of the upper conjunctiva area. There was no statistical difference between the two measurements of each index (P>0.05). Repeatability analysis results showed that coefficients of variation (CV) of all biological parameters of glands were less than 5% and the intraclass correlation coefficient (ICC) in both groups were greater than 0.95. Conclusion: The Meibomian Gland Bioimage Analyzer provides good reliability and consistency for morphological measurements of the meibomian gland, and it is expected to provide new non-invasive indicators for clinical assessment of the meibomian glands function.
