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

마우스 Retina OCT Fibergram Alignment with Confocal Images

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.

저자 스포트라이트: 녹내장 진단 및 치료 개선을 위한 In Vivo 및 Ex Vivo 망막 영상의 발전 

Align of Visible-Light Optical Coherence Tomography Fibergrams with Confocal Images of the Same Mouse Retina(가시광선 광간섭 단층 촬영 파이버그램과 동일한 마우스 망막의 컨포칼 이미지 정렬)

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  

In Vivo 및 Ex Vivo 마우스 망막 이미지의 정렬

먼저 각 마우스에서 얻은 4개의 이미지를 사용하여 합성 이미지를 만듭니다. 그래픽 편집기를 사용하여 모든 혈관을 정렬하고, 시신경 머리와 혈관 패턴을 랜드마크로 사용하여 복합 섬유 그램을 정렬하고, ICAM2로 염색된 동일한 망막의 합성 이미지 및 공초점 이미지를 겹쳐서 생체 내 및 생체 외 정렬을 달성합니다. 복합 vis-oct 섬유 그램은 평면 장착 망막의 해당 컨포칼 이미지와 비교됩니다.망막 신경절 축삭돌기에 대한 TUJ1을 사용한 면역 염색. 혈관은 공초점 이미지에서 ICAM2 표지된 혈관과 일치시킬 수 있는 구별 가능한 분지 구조를 나타냅니다. 생체 외 공초점 현미경과 in vivo vis-oct를 나란히 비교한 결과, 동일한 망막 신경절 축삭 다발 네트워크와 주변 망막 혈관 구조가 밝혀졌습니다.

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 ...