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

Allineamento del fibrogramma OCT della retina del topo con immagini confocali

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.

Autore in primo piano: Progressi nell'imaging retinico in vivo ed ex vivo per migliorare la diagnosi e il trattamento del glaucoma 

Allineamento di fibrogrammi per tomografia a coerenza ottica a luce visibile con immagini confocali della stessa retina murina

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  

Allineamento di immagini retiniche di topo in vivo ed ex vivo

Inizia creando un'immagine composita utilizzando le quattro immagini ottenute da ciascun mouse. Utilizzare un editor grafico per allineare tutti i vasi sanguigni, utilizzare la testa del nervo ottico e i modelli dei vasi sanguigni come punti di riferimento per l'allineamento del grammo di fibre composite sovrapporre le immagini composite e le immagini confocali della stessa retina colorata con ICAM2, per ottenere l'allineamento in vivo ed ex vivo. Il grammo di fibra vis-oct composito viene confrontato con la corrispondente immagine confocale della retina montata piatta.Immunocolorazione con TUJ1 per assoni RGC. I vasi sanguigni presentano strutture ramificate distinguibili che possono essere abbinate ai vasi sanguigni marcati ICAM2 sull'immagine confocale. Il confronto fianco a fianco tra microscopia confocale ex vivo e vis-oct in vivo ha rivelato reti di fasci assonali RGC identiche e vascolarizzazione retinica circostante.

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 ...

Ricerca

Neuroscienze

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

Tomografia a coerenza ottica (OCT) a luce visibile in vivo Imaging retinico di topo

Ex Vivo Confocal Imaging of Mouse Retina

Imaging confocale ex vivo della retina del topo