Goetz Martin, Memadathil Beena, Biesterfeld Stefan, Schneider Constantin, Gregor Sebastian, Galle Peter R, Neurath Markus F, Kiesslich Ralf
I. Medical Clinic, Johannes Gutenberg-Universitat, Mainz 55131, Germany.
World J Gastroenterol. 2007 Apr 21;13(15):2160-5. doi: 10.3748/wjg.v13.i15.2160.
To evaluate a newly developed hand-held confocal probe for in vivo microscopic imaging of the complete gastrointestinal tract in rodents.
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 microm multiply 475 microm. Optical slice thickness was 7 microm with a lateral resolution of 0.7 microm. Subsurface serial images at different depths (surface to 250 microm) were generated in real time at 1024 multiply 1024 pixels (0.8 frames/s) by placing the probe onto the tissue in gentle, stable contact. Tissue specimens were sampled for histopathological correlation.
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.
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.
评估一种新开发的手持式共聚焦探头，用于对啮齿动物的整个胃肠道进行体内显微成像。
设计了一种新型刚性共聚焦探头（直径7毫米），其光学特性类似于用于人体的柔性内镜显微镜系统，使用488纳米单线激光激发荧光团。在505至750纳米处检测光发射。视野为475微米×475微米。光学切片厚度为7微米，横向分辨率为0.7微米。通过将探头轻轻、稳定地接触组织，实时生成不同深度（表面至250微米）的地下序列图像，图像大小为1024×1024像素（0.8帧/秒）。采集组织标本进行组织病理学相关性分析。
在n = 48只小鼠体内以高分辨率观察到了食管、胃、小肠和大肠以及系膜、肝脏、胰腺和胆囊。使用共聚焦微型显微镜探头进行实时显微成像很容易实现。不同的染色方案（荧光素、吖啶黄、异硫氰酸荧光素标记的葡聚糖和番茄凝集素）各自突出了组织的特定方面，并且体内成像与传统组织学相关性极佳。体内血流监测为形态学成像增添了功能特性。
共聚焦显微镜在体内是可行的，能够以高分辨率观察整个胃肠道，甚至是地下组织结构。本研究评估的新型共聚焦探头设计与腹腔镜检查兼容，并显著扩展了可能应用于腹腔内器官的领域。它允许立即测试新的体内染色和应用选项，因此可以快速从动物研究转移到患者的临床应用。
