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. 2021 Aug 10;11:16252. doi: 10.1038/s41598-021-95320-z

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Confocal images of flat-mounted RPE from WT and Abca4^−/−mice. (a,c) Confocal images of the RPE cell mosaic acquired without emission filter. (b,d) Confocal images of RPE cell mosaic acquired with emission filter. (e,f) bar graphs showing the density of lipofuscin and melanosomes as number of granules per picoliter. The average number of lipofuscin granules and melanosomes were calculated from 9 confocal volumes per strain. White solid arrows on top panels indicate representative melanosomes, whereas the hollow arrows indicate representative lipofuscin granules. The red arrows indicate representative nuclei. White dashed-circles in (b,d) indicate the absesnce of melanosomes in the same region indicated by thick white arrows in (a,c). The melanosomes are not visible in (b,d) because the scattered light from the melanosomes is fully blocked by the fluorescense emission filter. The bottom panels in (a–d) show side views of the RPE volume. The dotted arrows in the bottom panel show the apical side of the RPE cells, whereas the solid arrows indicate the basal side of the RPE. (*p < 0.001,**p < 0.001). Scale bars: 25 μm.