Retinal Regeneration Following OCT-Guided Laser Injury in Zebrafish
Retinal Regeneration Following OCT-Guided Laser Injury in Zebrafish
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DOI：
10.1167/iovs.14-14724
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发表时间：
2014-10-01
影响因子：
4.4
通讯作者：
Yuan, Alex
中科院分区：
文献类型：
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作者：
DiCicco, Rose M.;Bell, Brent A.;Yuan, Alex
目的.采用光学相干断层扫描（OCT）引导的激光光凝技术建立斑马鱼局灶性损伤/再生模型。在室内空气中通过接触透镜通过OCT和共焦扫描激光检眼镜（cSLO）对成年斑马鱼成像。使用光束组合器，使用OCT C扫描图像进行靶向，应用532 nm激光光凝。将42至47 mW的激光光斑传递到视网膜。在损伤后的多个时间间隔，使用OCT和cSLO对鱼进行成像，以跟踪每个病变的进展。组织切片和TUNEL染色观察损伤反应。成功地应用了定位于外层视网膜的圆形损伤（26057 +/- 621 μ m（2））。激光应用通过实时OCT可视化，并且OCT和cSLO均可在体内检测到病变。病变后1天病变尺寸增加，然后尺寸减小。组织切片显示主要位于外层视网膜的损伤灶性区域。到3周时，受损区域已经再生，并重新建立了完全分层的结构。然而，OCT、cSLO成像和组织学仍然可以检测到细微变化。红外暗场成像在显示再生区域的细微变化方面比OCT更敏感。光学相干断层扫描引导的激光光凝是一种有用的工具，诱导局部病变和研究视网膜再生的斑马鱼。这种新的方法将使我们能够表征在正常和受损组织之间的界面处发生的细胞和分子变化。可以使用高分辨率OCT和cSLO成像在体内观察再生。
PURPOSE. Establish a focal injury/regeneration model in zebrafish using laser photocoagulation guided by optical coherence tomography (OCT).METHODS. Adult zebrafish were imaged by OCT and confocal scanning laser ophthalmoscopy (cSLO) in room air through a contact lens. Using a beam combiner, 532-nm laser photocoagulation was applied using the OCT C-scan image for targeting. Laser spots of 42 to 47 mW were delivered to the retina. At multiple intervals post injury, fish were imaged using both OCT and cSLO to follow the progression of each lesion. Histologic sections and TUNEL staining were performed to monitor the injury response.RESULTS. Round lesions (26057 +/- 621 mu m(2)) localized to the outer retina were successfully applied. Laser application was visualized by real-time OCT and lesions were detectable by both OCT and cSLO in vivo. Lesion size increased 1 day post lesion then decreased in size. Histologic sections showed focal areas of damage localized primarily to the outer retina. By 3 weeks, the damaged areas had regenerated and a fully laminated structure was re-established. However, subtle changes can still be detected by OCT, cSLO imaging, and histology. Infrared darkfield imaging was more sensitive than OCT at revealing subtle changes in regenerated areas.CONCLUSIONS. Optical coherence tomography-guided laser photocoagulation is a useful tool for inducing localized lesions and studying retinal regeneration in zebrafish. This novel method will allow us to characterize the cellular and molecular changes that take place at the interface between normal and damaged tissue. Regeneration can be observed using high-resolution OCT and cSLO imaging in vivo.