Purpose

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.

Methods

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.

Results

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.

Conclusions

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.

中文翻译：

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使用组织清除和光片显微镜对眼表面和完整的眼球进行高分辨率三维成像。

目的

眼表和完整眼球的高分辨率可视化对于我们理解和治疗眼疾至关重要。这项研究旨在使用先进的组织清除和三维（3D）成像技术来实现这一目标。

方法

研究中使用了Prox-1-GFP（绿色荧光蛋白）或Thy1-YFP（黄色荧光蛋白）的野生型和荧光标记的转基因小鼠。从正常或角膜炎症的疾病模型中收获眼球。将样品注入水凝胶单体并加热以进行聚合。通过电泳除去脂质。用高级光片荧光显微镜对前节或完整眼球的透明组织-水凝胶杂种进行免疫标记或内源性荧光成像。捕获了用于各种结构和细胞类型的高分辨率3D图像和视频。

结果

从正常和患病情况下完整的眼球可以实现光学透明性。通过其自然形态，位置和组织网络，可以检测到各种重要的结构和细胞类型，例如血液和淋巴管，Schlemm的管，神经和内皮细胞。

结论

这项研究使用组织清除和先进的光片显微镜技术，对完整的眼球进行了首次全面的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睛内部和外部在生物医学研究中的各种应用。