Figure 2 | Scientific Reports

Figure 2

From: Two-color in-resin CLEM of Epon-embedded cells using osmium resistant green and red fluorescent proteins

Figure 2

Two-color in-resin CLEM of nucleus and mitochondria was performed using H2B-CoGFPv0 and mCherry2-mito. Thin section (100 nm) of Epon-embedded cells expressing H2B-CoGFPv0 (green pseudo color) and mCherry2-mito (red pseudo color) was prepared. Fluorescent images (FM) were obtained in the presence of a TUK solution for multicolor with a BZ-X810 fluorescence microscope (CCD monochrome camera, NIKON CFI plan Apochromat 100 × Oil lens, gain + 16 dB, haze reduction) using filter sets for green and red fluorescent probes. Electron microscopic images (EM) were obtained with a Helios NanoLab 660 scanning electron microscope (a backscattered electron detector at a voltage of 2.0 kV with a current of 0.4 nA), and were processed with a method called “Contrast Limited Adaptive Histogram Equalization” using imageJ software with the plugin Enhance Local Contrast (CLAHE). The “Merge” is a merged image of the fluorescence image (FM) with an electron microscopic image (EM). The images in (B) indicate magnification of images corresponding to the boxed area in the Merge image in (A).

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