非分离小鼠脉络膜/视网膜色素上皮/视网膜染色及整装片用于视网膜下层面的整体观察
Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer.
作者信息
Lee Sung-Jin, Kim Soo-Young
机构信息
R&D Center II, ExosomePlus, Inc., Seoul St. Mary's Hospital 222, Banpo-daero, Seocho-gu, Seoul, South Korea.
出版信息
Bio Protoc. 2021 Jan 5;11(1):e3872. doi: 10.21769/BioProtoc.3872.
The subretinal layer between retinal pigment epithelium (RPE) and photoreceptors is a region involved in inflammation and angiogenesis during the procession of diseases such as age-related macular degeneration. The current protocols of whole mounts (retina and RPE) are unable to address the intact view of the subretinal layer because the separation between retina and RPE is required, and each separate tissue is then stained. Non-separate Sclerochoroid/RPE/Retina whole mount staining was recently developed and reported. The method can be further combined and optimized with melanin bleaching and tissue clearing. Here, we describe steps of both non-pigmented and pigmented mouse Sclerochoroid/RPE/Retina whole mount including eyeball preparation, staining, mounting and confocal scanning. In addition, we present the confocal images of RPE, subretinal microglia and the neighboring photoreceptors in Sclerochoroid/RPE/Retina whole mounts.
视网膜色素上皮（RPE）和光感受器之间的视网膜下层是年龄相关性黄斑变性等疾病进程中涉及炎症和血管生成的区域。目前的全层装片（视网膜和RPE）方案无法呈现视网膜下层的完整视图，因为需要将视网膜和RPE分离，然后对每个单独的组织进行染色。最近开发并报道了非分离的巩膜脉络膜/RPE/视网膜全层染色方法。该方法可进一步与黑色素漂白和组织透明化相结合并优化。在此，我们描述了非色素和色素小鼠巩膜脉络膜/RPE/视网膜全层装片的步骤，包括眼球制备、染色、封片和共聚焦扫描。此外，我们展示了巩膜脉络膜/RPE/视网膜全层装片中RPE、视网膜下小胶质细胞和相邻光感受器的共聚焦图像。
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