Figure 2
From: Hydrogel-based milliwell arrays for standardized and scalable retinal organoid cultures
(A) Schematic representation and photograph of histological sectioning of ROAs. The entire gel array containing the organoids is embedded in OCT and, (B), frozen in order to be cryo-sectioned. (C) Representative wide-field fluorescence tile scan of an entire section of a ROA. Green fluorescence represents the GFP expressed in photoreceptor cells. The dotted line represents the milliwell edges. (D–J) Immunocytochemistry of Crx-GFP-derived retinal organoids cultured on milliwell arrays to detect the retinal cell type composition at day 26 of differentiation. (Di–Ji) GFP-positive cells represent the photoreceptors. (Dii) PAX6-positive ganglion and amacrine cells, (Eii) CALBINDING-positve horizonatl cells, (Fii) CRALBP-positive Muller glia, (Gii) OTX2-positive, CRX-negative bipolar cells, (Hii) GNAT2-positive cone cells, (Iii) GNAT1-positive rod photoreceptors in red and, (Jii), Rhodopsin-positive photoreceptors in red, their respective merged counterparts (Diii–Jiii). White dashed lines indicate the partially occurred lamination process to separate the neuroblastic layer, where GFP-positive photoreceptor localised, from the underneath ganglion cell layer. Scale bar: 100 μm.
