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. Author manuscript; available in PMC: 2013 Dec 24.
Published in final edited form as: Methods Cell Biol. 2012;111:10.1016/B978-0-12-416026-2.00015-7. doi: 10.1016/B978-0-12-416026-2.00015-7

Fig. 1.

Fig. 1

Schematic diagram showing the procedure for nano-fEM. Transgenic animals (A) are subject to high-pressure freezing (B, C). The cellular water is substituted with acetone while tissues are fixed with 0.1% potassium permanganate and 0.001% osmium tetroxide (D, E). The specimens are embedded into GMA plastic (F). The plastic block is trimmed to the region of interest (G), and thin slices (~80 nm thick) of tissues are cut using a diamond knife (H). Sections are imaged first with super-resolution fluorescence microscopy (I) and second with scanning electron microscopy (J). The correlative image is then aligned based on the fiducial markers (K).