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. 2019 Apr 28;8(5):391. doi: 10.3390/cells8050391

Figure 2.

Figure 2

PACT greatly improves retinal organoid immunocytochemistry and high-resolution imaging. Retinal organoids were immunostained and imaged by light sheet microscopy. Single planes were selected from an acquired image stack to visualize differences between uncleared (left panel) and cleared (right panel) retinal organoids. Each panel contains an overview (left image) and magnified cutout (right image). Organoids were labeled as followed: (a) day 182 retinal organoids were labeled with DAPI (4′,6-Diamidin-2-phenylindol, nuclei staining, yellow). Second and forth image show a magnified area as indicated in the first and third image, respectively. (b) Day 182 retinal organoids were stained for Recoverin (pan-photoreceptor marker). Second and forth image show a magnified area as indicated in the first and third image, respectively. (c) Day 182 retinal organoids were previously transfected with a lentiviral vector labeling photoreceptor cells (pJG-IRPB-eGFP). Cleared or uncleared retinal organoids were additionally stained by a GFP-antibody. (d) Day 290 retinal organoids were stained for PKCα (bipolar cell marker, light blue) and DAPI (nuclei, white). Second and forth image show xy-, xz-, and yz-plane views of a magnified bipolar cell. Scale bars: (a–c) 100 µm, (d) 50 µm.