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

Alinhamento de Fibra de OCT de Retina de Camundongo com Imagens Confocais

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 em destaque: Avanços na imagem da retina in vivo e ex vivo para melhorar o diagnóstico e o tratamento do glaucoma 

Alinhamento de Fibrogramas de Tomografia de Coerência Óptica com Luz Visível com Imagens Confocais da Mesma Retina de Camundongo

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  

Alinhamento de Imagens da Retina de Camundongos In Vivo e Ex Vivo

Comece criando uma imagem composta usando as quatro imagens obtidas de cada mouse. Utilize um editor gráfico para alinhar todos os vasos sanguíneos, empregue os padrões da cabeça do nervo óptico e dos vasos sanguíneos como pontos de referência para alinhar a sobreposição de grama da fibra composta, as imagens compostas e as imagens confocais da mesma retina coradas com ICAM2, para alcançar o alinhamento in vivo e ex vivo. O grama de fibra vis-oct composto é comparado com a imagem confocal correspondente da retina plana montada.Imunocoloração com TUJ1 para axônios RGC. Os vasos sanguíneos exibem estruturas ramificadas distinguíveis que podem ser combinadas com os vasos sanguíneos marcados com ICAM2 na imagem confocal. A comparação lado a lado entre a microscopia confocal ex vivo e a vis-oct in vivo revelou redes idênticas de feixes de axônios RGC e vasculatura retiniana circundante.

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 ...