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. 2021 Dec 15;9:737621. doi: 10.3389/fcell.2021.737621

FIGURE 2.

FIGURE 2

Fluorescence and electron micrographs of mouse hippocampus tissue; IFR protocol freeze-substituted with 0.2% uranyl acetate. The fluorescent signal was preserved in a 100 µm hippocampus slice after IRF protocol as observed in the resin block imaged with widefield Zeiss Axioplan 2 with Plan-NeoFluar 63x lens. (A) and after thin sectioning imaged with a Nikon TiE inverted C2 confocal microscope with a Plan Apo VC 20x dry lens (B, right) and Plan Apo 10x dry lens (B, left). An enlarged view of the area enclosed in the black square in the left image of (B) shows pyramidal cells in CA1 (B, right). (C, D) TEM imaging of GFP positive CA1 hippocampal cells, showing good ultrastructure. (D) A higher magnification image of the area indicated by the black box in C. (E, F) same sample block as in (C, D), but imaged by SEM using a BSE detector (Gatan OnPoint detector) at 1 kV. The image quality (inverse contrast) is comparable to TEM imaging. WD = 6 mm. Section thickness = 100 nm. Abbreviations: BSE, backscattered electrons detector; GFP, green fluorescent protein; IRF, in-resin fluorescence; SEM, scanning electron microscope; TEM, transmission electron microscopy; UrAc, uranyl acetate; WD, working distance. Scale bars: A, B left = 100 μm; B, right = 40 μm; C, E = 1 μm; D, F = 0.5 µm.