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

Alignement du fibergramme OCT de la rétine de la souris avec des images confocales

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.

Pleins feux sur l’auteur : Progrès de l’imagerie rétinienne in vivo et ex vivo pour améliorer le diagnostic et le traitement du glaucome 

Alignement de fibrogrammes de tomographie par cohérence optique en lumière visible avec des images confocales de la même rétine de souris

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  

Alignement d’images rétiniennes de souris in vivo et ex vivo

Commencez par créer une image composite à l’aide des quatre images obtenues à partir de chaque souris. Utilisez un éditeur graphique pour aligner tous les vaisseaux sanguins, utilisez la tête du nerf optique et les modèles de vaisseaux sanguins comme points de repère pour aligner le gramme de fibre composite chevaucher les images composites et les images confococaux de la même rétine colorée avec ICAM2, pour obtenir un alignement in vivo et ex vivo. Le gramme de fibre vis-oct composite est comparé à l’image confocale correspondante de la rétine montée à plat.Immuno coloration avec TUJ1 pour les axones RGC. Les vaisseaux sanguins présentent des structures ramifiées distinctes qui peuvent être appariées aux vaisseaux sanguins marqués ICAM2 sur l’image confocale. La comparaison côte à côte entre la microscopie confocale ex vivo et la vis-oct in vivo a révélé des réseaux de faisceaux axonaux RGC identiques et une vascularisation rétinienne environnante.

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 ...

Recherche

Neurosciences

In Vivo Visible Light Optical Coherence Tomography (OCT) Mouse Retinal Imaging

Tomographie par cohérence optique (OCT) en lumière visible in vivo Imagerie rétinienne de souris

Ex Vivo Confocal Imaging of Mouse Retina

Imagerie confocale ex vivo de la rétine de souris