Timing: 1 h

Use FEI Talos L120C TEM equipped with CETA 16 MP camera or a TEM with similar configuration to obtain images of ultrathin sections (Figures 4B and 4C).

Mount the grid in a standard TEM holder. Place the specimen facing down inside the specimen carrier. Then insert the holder into the TEM (Figure 4A).

Turn the magnification down to find the stained sheets. Scan the grid at low magnification (×1250) to get an overall image of the myelinated regions. The acceleration voltage is 80 kV.

The Stage2 control panel provides functionality for storing positions and tracks (Figure 4D). We firstly use the Stage2 control panel to label the outline of the myelinated region of a tissue. Then we randomly acquire 5–30 images per section (5 images for optic nerve and corticospinal tract, 30 images for corpus callosum) at higher magnification. To avoid taking repeated images, we use the Stage2 control panel to mark the stage position of different images.

Remove the grid from the TEM.

Illustration of TEM image taken

(A) Mount the grid in a standard TEM holder.

(B and C) FEI Talos L120C TEM equipped with CETA 16 MP camera to obtain images.

(D) Stage2 Control Panel provides functionality for controlling the CompuStage, storing positions and tracks.

Wang Y., Sun B., Shibata B, & Guo F. (2022). Transmission electron microscopic analysis of myelination in the murine central nervous system. STAR Protocols, 3(2), 101304.
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