alignment of in vivo and ex vivo mouse retinal images

小鼠视网膜 OCT 纤维图与共聚焦图像的比对

Retinal ganglion cells (RGCs) play a critical role in visual information processing, receiving synaptic inputs through their dendritic trees in the inner plexiform layer (IPL) and transmitting the information via their axons in the retinal nerve fiber layer (RNFL) to the brain1,2,3,4. In diseased conditions such as glaucoma, early RGC degeneration may result in subtle changes in the RNFL, the ganglion cell layer (GCL), the IPL, and the optic nerve in both patients and rodent models5,6,7,8,9. Early detection of these morphological changes in RGCs is thus essential for timely intervention to prevent RGC and vision loss.
We have recently developed a new clinical-ready imaging technology called visible-light optical coherence tomography (vis-OCT) to satisfy the need for in vivo monitoring of RGC damage. Vis-OCT improved the axial resolution, reaching 1.3 &#181;m in the retina10, 11, allowing for the visualization of individual RGC axon bundles in the RNFL. Subsequently, vis-OCT fibergraphy (vis-OCTF) was established to track and quantify RGC damage at the single axon bundle level in mice11,12,13. However, ex vivo confocal imaging of the same retina as the gold standard is often necessary to validate the in vivo findings. Therefore, this study will demonstrate how to align in vivo images acquired by vis-OCTF with ex vivo confocal images of the same mouse retina. The protocol aims to validate the in vivo findings by ex vivo confocal imaging and establish a foundation for examining the molecular and cellular changes underlying RGC damage in diseased conditions.

作者聚焦：用于改善青光眼诊断和治疗的体内和离体视网膜成像的进展 

可见光光学相干断层扫描光纤图与同一小鼠视网膜的共聚焦图像的对齐

Watch this Scientific Journal Video about Alignment of In Vivo and Ex Vivo Mouse Retinal Images at JoVE.com

Alignment of In Vivo and Ex Vivo Mouse Retinal Images  

体内和离体小鼠视网膜图像的比对

首先使用从每只鼠标获得的四个图像创建复合图像。利用图形编辑器对齐所有血管，以视神经头和血管图案为标志，对齐复合纤维革兰重叠的复合图像和ICAM2染色的同一视网膜的共聚焦图像，实现体内和离体对齐。将复合的 vis-oct 纤维克与相应的平面视网膜共聚焦图像进行比较。使用 TUJ1 对 RGC 轴突进行免疫染色。血管表现出可区分的分支结构，可以与共聚焦图像上的 ICAM2 标记的血管相匹配。离体共聚焦显微镜和体内vs-oct之间的并排比较显示相同的RGC轴突束网络和周围的视网膜血管系统。

alignment-of-in-vivo-and-ex-vivo-mouse-retinal-images

Research

JoVE Journal

Neuroscience

Alignment of In Vivo and Ex Vivo Mouse Retinal Images

132 次观看. 首先使用从每只鼠标获得的四个图像创建复合图像。利用图形编辑器对齐所有血管，以视神经头和血管图案为标志，对齐复合纤维革兰重叠的复合图像和ICAM2染色的同一视网膜的共聚焦图像，实现体内和离体对齐。将复合的 vis-oct 纤维克与相应的平面视网膜共聚焦图像进行比较。使用 TUJ1 对 RGC 轴突进行免疫染色。血管表现出可区分的分支结构，可以与共聚焦图像上的 IC...

视频: Alignment of In Vivo and Ex Vivo Mouse Retinal Images  

Alignment of Visible-Light Optical Coherence Tomography Fibergrams with Confocal Images of the Same Mouse Retina

In recent years, in vivo retinal imaging, which provides non-invasive, real-time, and longitudinal information about biological systems and processes, has been increasingly applied to obtain an objective assessment of neural damage in eye diseases. Ex vivo confocal imaging of the same retina is often necessary to validate the in vivo findings especially in animal research. In this study, we demonstrated a method for aligning an ex vivo confocal image of the mouse retina with its in vivo images. A new clinical-ready imaging technology called visible light optical coherence tomography fibergraphy (vis-OCTF) was applied to acquire in vivo images of the mouse retina. We then performed the confocal imaging of the same retina as the &#34;gold standard&#34; to validate the in vivo vis-OCTF images. This study not only enables further investigation of the molecular and cellular mechanisms but also establishes a foundation for a sensitive and objective evaluation of neural damage in vivo.

The present protocol outlines the steps for aligning in vivo visible-light optical coherence tomography fibergraphy (vis-OCTF) images with ex vivo confocal images of the same mouse retina for the purpose of verifying the observed retinal ganglion cell axon bundle morphology in the in vivo images.

In recent years, in vivo retinal imaging, which provides non-invasive, real-time, and longitudinal information about biological systems and processes, has ...