alignment of in vivo and ex vivo mouse retinal images

Alineación de Fibergram OCT de Retina de Ratón con Imágenes 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.

Autor destacado: Avances en imágenes de retina in vivo y ex vivo para mejorar el diagnóstico y tratamiento del glaucoma 

Alineación de figramas de tomografía de coherencia óptica en luz visible con imágenes confocales de la misma retina de ratón

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  

Alineación de imágenes retinianas de ratón in vivo y ex vivo

Comience creando una imagen compuesta utilizando las cuatro imágenes obtenidas de cada ratón. Utilice un editor de gráficos para alinear todos los vasos sanguíneos, emplee la cabeza del nervio óptico y los patrones de los vasos sanguíneos como puntos de referencia para alinear la fibra compuesta, superponga las imágenes compuestas y las imágenes confocales de la misma retina teñida con ICAM2, para lograr la alineación in vivo y ex vivo. El gramo de fibra vis-oct compuesto se compara con la imagen confocal correspondiente de la retina montada en plano.Inmunotinción con TUJ1 para axones de RGC. Los vasos sanguíneos exhiben estructuras ramificadas distinguibles que pueden coincidir con los vasos sanguíneos marcados con ICAM2 en la imagen confocal. La comparación lado a lado entre la microscopía confocal ex vivo y la vis-oct in vivo reveló redes idénticas de haces de axones RGC y vasculatura retiniana circundante.

alignment-of-in-vivo-and-ex-vivo-mouse-retinal-images

Research

JoVE Journal

Neuroscience

Alignment of In Vivo and Ex Vivo Mouse Retinal Images

132 vistas. Comience creando una imagen compuesta utilizando las cuatro imágenes obtenidas de cada ratón. Utilice un editor de gráficos para alinear todos los vasos sanguíneos, emplee la cabeza del nervio óptico y los patrones de los vasos sanguíneos como puntos de referencia para alinear la fibra compuesta, superponga las imágenes compuestas y las imágenes confocales de la misma retina teñida con ICAM2, para lograr la alineación in vivo y ex vivo. El gramo de fibra vis-oct compuesto se compara...

Video: 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 been increasingly applied to obtain an objective assessment of neural damage in eye diseases. Ex vivo confocal imaging of the same retina is often necessary to validate the in vivo findings especially in animal research. In this study, we demonstrated a method for aligning an ex vivo confocal image of the mouse retina with its in vivo images. A new clinical-ready imaging technology called visible light optical coherence tomography fibergraphy (vis-OCTF) was applied to acquire in vivo images of the mouse retina. We then performed the confocal imaging of the same retina as the &#34;gold standard&#34; to validate the in vivo vis-OCTF images. This study not only enables further investigation of the molecular and cellular mechanisms but also establishes a foundation for a sensitive and objective evaluation of neural damage in vivo.

The present protocol outlines the steps for aligning in vivo visible-light optical coherence tomography fibergraphy (vis-OCTF) images with ex vivo confocal images of the same mouse retina for the purpose of verifying the observed retinal ganglion cell axon bundle morphology in the in vivo images.

In recent years, in vivo retinal imaging, which provides non-invasive, real-time, and longitudinal information about biological systems and processes, has ...