Name: Alignment of In Vivo and Ex Vivo Mouse Retinal Images
Uploaded: 2023-06-30T13:00:00
Description: 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

マウス Retina OCT ファイバーグラムと共焦点画像によるアライメント

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.

著者スポットライト:緑内障の診断と治療を改善するためのIn vivoおよびex vivo網膜イメージングの進歩 

可視光光干渉断層撮影ファイバーグラムと同一マウス網膜の共焦点像のアライメント

We present a protocol for aligning in vivo VIS-OCT images with ex vivo immunostained retinal nerve fibers to validate our novel imaging technology. VIS-OCT enables visualization of retinal nerve fibers in vivo, and this process confirms the accuracy of our findings. Our protocol combines in vivo and ex vivo imaging to provide a real time visualization of retina structure and confirm findings.The process is applicable to mouse models of eye diseases, provided the anterior eye portion allows for optical imaging. This includes diabetic retinopathy and retinal ischemia models. These studies lay the foundation for establishing an objective evaluation of neuro damage in humans, which can significantly improve glaucoma diagnosis and treatment in the future.

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  

In vivoおよびEx Vivoマウス網膜画像のアライメント

まず、各マウスから取得した 4 つの画像を使用して合成画像を作成します。グラフィックエディタを利用してすべての血管をアライメントし、視神経乳頭と血管パターンをランドマークとして使用して、複合ファイバーグラムをアライメントし、ICAM2で染色された同じ網膜の合成画像と共焦点画像を重ねて、in vivoおよびex vivoアライメントを実現します。複合可視線維グラムは、フラットマウント網膜の対応する共焦点画像と比較されます。RGC軸索のTUJ1による免疫染色。血管は、共焦点画像上のICAM2標識血管と一致する識別可能な分岐構造を示します。ex vivo 共焦点顕微鏡と in vivo vis-oct を並べて比較すると、同一の RGC 軸索バンドル ネットワークと周囲の網膜血管系が明らかになりました。

Research

JoVE Journal

Neuroscience

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 ...