High resolution three-dimensional imaging of the ocular surface and intact eyeball using tissue clearing and light sheet microscopy.
High resolution three-dimensional imaging of the ocular surface and intact eyeball using tissue clearing and light sheet microscopy.
复制标题
DOI：
10.1016/j.jtos.2020.04.009
复制
发表时间：
2020-07
期刊：
影响因子：
--
通讯作者：
Chen L
中科院分区：
文献类型：
--
作者：
Yang Y;Li G;Chen L
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科疾病至关重要。本研究旨在使用先进的组织清除和三维（3D）成像技术实现这一目标。在研究中使用野生型和荧光标记的Prox-1-GFP（绿色荧光蛋白）或Thy 1-YFP（黄色荧光蛋白）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聚合。通过电泳除去脂质。通过先进的光片荧光显微镜对具有免疫标记或内源性荧光的前节或完整眼球的透明组织-水凝胶杂交体进行成像。高分辨率的3D图像和视频被捕获的结构和细胞类型的广泛阵列。从正常和患病条件下的完整眼球获得光学透明度。各种重要的结构和细胞类型，如血管和淋巴管，Schlemm管，神经和内皮细胞，检测到它们的自然形态，位置和组织网络。这项研究提供了第一个全面的和三维高分辨率成像的完整眼球使用组织清除和先进的光片显微镜。鉴于眼睛是身体的窗口，我们预计这项先进的技术将促进眼睛内外生物医学研究的各种应用。
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 labelled 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.
登录
查看更多内容
影响因子：
82.9
作者：
Chen, L;Hamrah, P;Dana, MR
通讯作者：
Dana, MR
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.7
作者：
Truong T;Boshra H;Embury-Hyatt C;Nfon C;Gerdts V;Tikoo S;Babiuk LA;Kara P;Chetty T;Mather A;Wallace DB;Babiuk S
通讯作者：
Babiuk S
影响因子：
4.6
作者：
Kuwajima, Takaaki;Sitko, Austen A.;Mason, Carol
通讯作者：
Mason, Carol
影响因子：
3.4
作者：
Henning, Yoshiyuki;Osadnik, Christin;Malkemper, Erich Pascal
通讯作者：
Malkemper, Erich Pascal