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

Muis Retina OCT Fibergram-uitlijning met confocale beelden

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.

Auteur in de schijnwerpers: vooruitgang in in vivo en ex vivo retinale beeldvorming voor verbeterde diagnose en behandeling van glaucoom 

Uitlijning van optische coherentietomografievezelgrammen in zichtbaar licht met confocale beelden van hetzelfde netvlies van de muis

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  

Uitlijning van in vivo en ex vivo netvliesbeelden van muizen

Begin met het maken van een samengestelde afbeelding met behulp van de vier verkregen afbeeldingen van elke muis. Gebruik een grafische editor om alle bloedvaten uit te lijnen, gebruik de oogzenuwkop en bloedvatpatronen als oriëntatiepunten voor het uitlijnen van de composietvezelgram, overlap de samengestelde afbeeldingen en de confocale afbeeldingen van hetzelfde netvlies gekleurd met ICAM2, om in vivo en ex vivo uitlijning te bereiken. De samengestelde vis-oct vezelgram wordt vergeleken met het corresponderende confocale beeld van plat gemonteerd netvlies.Immunokleuring met TUJ1 voor RGC-axonen. Bloedvaten vertonen te onderscheiden vertakkingsstructuren die kunnen worden vergeleken met de ICAM2-gelabelde bloedvaten op het confocale beeld. Zij-aan-zij vergelijking tussen ex vivo confocale microscopie en in vivo vis-oct onthulde identieke RGC Axonenbundelnetwerken en omringende retinale vasculatuur.

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 ...