Samples preparation, imaging for TEM and phagosomes analysis were performed as reported previously
32 (link)
. Briefly, zebrafish samples were prepared for TEM using the same protocol for light microscopy. 80 nm sections were cut on a Leica EM UC6 microtome and mounted on copper grids and post-stained with 2% uranyl acetate and 3% lead citrate. The optic nerve was used as a reference point for sectioning. Imaging was performed on an FEI Tecnai 120 electron microscope. By TEM, phagosomes were manually counted, and the density was calculated as phagosomes per micron of RPE. A minimum of 350 μm of RPE surface was analysed per larva.
32 (link)
. Briefly, zebrafish samples were prepared for TEM using the same protocol for light microscopy. 80 nm sections were cut on a Leica EM UC6 microtome and mounted on copper grids and post-stained with 2% uranyl acetate and 3% lead citrate. The optic nerve was used as a reference point for sectioning. Imaging was performed on an FEI Tecnai 120 electron microscope. By TEM, phagosomes were manually counted, and the density was calculated as phagosomes per micron of RPE. A minimum of 350 μm of RPE surface was analysed per larva.
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