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首页> 外文期刊>World Journal of Gastroenterology >In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy
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In vivo subsurface morphological and functional cellular and subcellular imaging of the gastrointestinal tract with confocal mini-microscopy

机译：共聚焦微型显微镜对胃肠道的体内亚表面形态学和功能性细胞和亚细胞成像

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AIM: To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents. METHODS: A novel rigid confocal probe (diameter 7 mm) was designed with optical features similar to the flexible endomicroscopy system for use in humans using a 488 nm single line laser for fluorophore excitation. Light emission was detected at 505 to 750 nm. The field of view was 475 μm x 475 μm. Optical slice thickness was 7 μm with a lateral resolution of 0.7 μm. Subsurface serial images at different depths (surface to 250 μm) were generated in real time at 1024 x 1024 pixels (0.8 frames/s) by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation. RESULTS: The esophagus, stomach, small and large intestine and meso, liver, pancreas and gall bladder were visualised in vivo at high resolution in n = 48 mice. Real time microscopic imaging with the confocal mini-microscopy probe was easy to achieve. The different staining protocols (fluorescein, acriflavine, FITC-labelled dextran and L. esculentum lectin) each highlighted specific aspects of the tissue, and in vivo imaging correlated excellently with conventional histology. In vivo blood flow monitoring added a functional quality to morphologic imaging. CONCLUSION: Confocal microscopy is feasible in vivo allowing the visualisation of the complete GI tract at high resolution even of subsurface tissue structures. The new confocal probe design evaluated in this study is compatible with laparoscopy and significantly expands the field of possible applications to intra-abdominal organs. It allows immediate testing of new in vivo staining and application options and therefore permits rapid transfer from animal studies to clinical use in patients.
机译：目的：评估一种新开发的手持共聚焦探针，用于对啮齿类动物的整个胃肠道进行体内显微成像。方法：设计了一种新颖的刚性共聚焦探针（直径7 mm），其光学特征类似于柔性内窥镜系统，适用于使用488 nm单线激光器进行荧光团激发的人体。在505至750nm处检测到发光。视场为475μmx 475μm。光学切片厚度为7μm，横向分辨率为0.7μm。通过将探针以柔和，稳定的接触方式放置在组织上，可以实时生成1024 x 1024像素（0.8帧/秒）的不同深度（表面至250μm）的地下串行图像。取样组织标本进行组织病理学相关性分析。结果：在n = 48只小鼠体内，可以高分辨率观察到食道，胃，小肠和大肠以及中观，肝脏，胰腺和胆囊。使用共聚焦微型显微镜探针进行实时显微成像很容易实现。不同的染色方案（荧光素，a啶黄，FITC标记的右旋糖酐和食用植物凝集素）均突出了组织的特定方面，并且体内成像与常规组织学密切相关。体内血流监测为形态成像增加了功能质量。结论：共聚焦显微镜在体内是可行的，即使在表面下的组织结构下，也可以高分辨率显示完整的胃肠道。在这项研究中评估的新的共聚焦探头设计与腹腔镜检查兼容，并且极大地扩展了可能在腹腔内器官中应用的领域。它允许立即测试新的体内染色和应用选项，因此可以从动物研究快速转移到患者的临床使用。

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