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

Mus Retina OCT Fibergram Justering med Konfokale Billeder

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.

Forfatter Spotlight: Fremskridt inden for in vivo og ex vivo retinal billeddannelse til forbedret glaukomdiagnose og behandling 

Justering af optisk kohærenstomografi i synligt lys med konfokale billeder af samme musenethinde

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  

Justering af in vivo og ex vivo mus retinale billeder

Start med at oprette et sammensat billede ved hjælp af de fire opnåede billeder fra hver mus. Brug en grafikeditor til at justere alle blodkarrene, anvende synsnervehovedet og blodkarmønstre som landemærker til justering af det sammensatte fibergramoverlap de sammensatte billeder og de konfokale billeder af den samme nethinden farvet med ICAM2 for at opnå in vivo og ex vivo justering. Det sammensatte vis-oktfibergram sammenlignes med det tilsvarende konfokale billede af fladmonteret nethinden.Immun plet med TUJ1 til RGC axoner. Blodkar udviser skelnelige forgreningsstrukturer, som kan matches med ICAM2-mærkede blodkar på det konfokale billede. Side-by-side sammenligning mellem ex vivo konfokal mikroskopi og in vivo vis-oct afslørede identiske RGC Axon bundle netværk og omgivende retinal vaskulatur.

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 ...