Non-separate Mouse Sclerochoroid/RPE/Retina Staining and Whole Mount for the Integral Observation of Subretinal Layer.

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.