Vision Science Graduate Program, Center for Eye Disease and Development, and School of Optometry, University of California, Berkeley, CA, 94720, USA; Department of Ophthalmology, The Second Xiangya Hospital, Central South University, Changsha, 410011, China.
Vision Science Graduate Program, Center for Eye Disease and Development, and School of Optometry, University of California, Berkeley, CA, 94720, USA.
Ocul Surf. 2020 Jul;18(3):526-532. doi: 10.1016/j.jtos.2020.04.009. Epub 2020 May 14.
High resolution visualization of the ocular surface and intact eyeball is critical and essential for our understanding and treatment of eye diseases. This study is to achieve this goal using advanced tissue clearing and three-dimensional (3D) imaging technologies.
Wild type and fluorescently labeled transgenic mice of Prox-1-GFP (green fluorescent protein) or Thy1-YFP (yellow fluorescent protein) were used in the study. Eyeballs were harvested from normal or a disease model of corneal inflammation. Samples were infused with hydrogel monomers and heated for polymerization. Lipids were removed by electrophoresis. The transparent tissue-hydrogel hybrids of the anterior segments or intact eyeballs with immunolabeling or endogenous fluorescence were imaged by an advanced light sheet fluorescent microscope. High resolution 3D images and videos were captured for a wide array of structures and cell types.
Optical transparency was achieved from intact eyeballs of both normal and diseased conditions. A variety of important structures and cell types, such as blood and lymphatic vessels, Schlemm's canal, nerves and endothelial cells, were detected with their natural morphology, location and organizational network.
This study provides the first comprehensive and 3D high resolution imaging of the intact eyeball using tissue clearing and advanced light sheet microscopy. Given that the eye is the window of the body, we anticipate this advanced technology will facilitate diverse applications in biomedical research inside and outside the eye.
高分辨率可视化眼部表面和完整眼球对于我们理解和治疗眼部疾病至关重要。本研究旨在使用先进的组织透明化和三维（3D）成像技术来实现这一目标。
本研究使用了 Prox-1-GFP（绿色荧光蛋白）或 Thy1-YFP（黄色荧光蛋白）荧光标记的野生型和转基因小鼠。从正常或角膜炎症疾病模型中采集眼球。将样品注入水凝胶单体并加热聚合。通过电泳去除脂质。对具有免疫标记或内源性荧光的前节透明组织-水凝胶混合物或完整眼球进行先进的光片荧光显微镜成像。捕获了广泛的结构和细胞类型的高分辨率 3D 图像和视频。
从正常和患病条件下的完整眼球中实现了光学透明度。检测到各种重要的结构和细胞类型，如血管和淋巴管、施莱姆氏管、神经和内皮细胞，具有其自然形态、位置和组织网络。
本研究首次使用组织透明化和先进的光片显微镜对完整眼球进行了全面的 3D 高分辨率成像。鉴于眼睛是身体的窗口，我们预计这项先进技术将促进眼内和眼外生物医学研究中的多种应用。
