Choi Myunghwan, Yun Seok Hyun
Opt Express. 2013 Dec 16;21(25):30842-8. doi: 10.1364/OE.21.030842.
Regeneration of the intestinal epithelium after injury or during pathogenesis is a dynamic cellular process critical for host immunity. However, current epithelial injury models provide poor spatial control, complicating the study of precise cellular responses. Here we developed endoscopic femtosecond-laser surgery capable of generating acute tissue injury. A side-view probe provides a convenient access to the distal colon in the mouse in vivo and allows real-time intraoperative monitoring as well as pre- and post-surgery examinations via multiphoton imaging. The photo-induced damage showed a nonlinear dependence on laser intensity. At an optical power of 200 mW (2.5 nJ per pulse), scanning the beam focus over 300x300 µm(2) area in the colonic mucosa generated substantial vascular damages within 30 s. We confirmed the localized tissue damage and the physiologic regeneration of the disrupted epithelium by in situ barrier function assays, validating the animal model for epithelial regeneration following injury. The femtosecond endosurgery technique is applicable to various experimental models based on laser-induced perturbations.
损伤后或发病过程中肠上皮的再生是一个对宿主免疫至关重要的动态细胞过程。然而，目前的上皮损伤模型在空间控制方面较差，使得精确细胞反应的研究变得复杂。在此，我们开发了能够产生急性组织损伤的内镜飞秒激光手术。一个侧视探头可方便地进入小鼠体内的远端结肠，并允许通过多光子成像进行实时术中监测以及手术前后检查。光诱导损伤显示出对激光强度的非线性依赖性。在200 mW的光功率（每脉冲2.5 nJ）下，将光束焦点扫描结肠黏膜上300×300 µm²的区域，在30秒内产生了大量血管损伤。我们通过原位屏障功能测定证实了局部组织损伤以及受损上皮的生理再生，验证了损伤后上皮再生的动物模型。飞秒内镜手术技术适用于基于激光诱导扰动的各种实验模型。
