Use of three-dimensional electron tomography to distinguish different regions of the endomembrane system. Use of three-dimensional electron tomography.

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Use of three-dimensional electron tomography to distinguish different regions of the endomembrane system. Use of three-dimensional electron tomography to distinguish different regions of the endomembrane system. (A) Transmission electron microscopy (TEM) image of a prepared tomographic slice (4.3 mm thick) through a region of the Golgi complex and surrounding cytoplasm. Take note of the division of the Golgi stack from cis-Golgi (cis) to medial-Golgi (med) to trans-Golgi (trans) and the trans-Golgi network (TGN). Specialized labeling techniques can be used to help distinguish different cisternae. A non-coated vesicle (NCV) and a multivesicular body (MVB) are also labeled. (B) Here, cis-, medial- and trans-Golgi cisternae seen in A are visualized as a three-dimensional image, displaying the divisions of the Golgi stack, as well as a distinct TGN with associated non-coated and clathrin-coated vesicles (CCVs). MVBs are also clearly visible as white bodies attached to the TGN. (C) Top-down or face-on views of the Golgi stack. 3D electron tomography allows for the virtual dissection of the Golgi stack, from top to bottom (from top, cisternae are indicated: cis1, cis 2, medial 1, medial 2, medial 3, trans 2, TGN). The colors for each cisterna are as in B. This top-down view also allows visualization of the formation of protein aggregates (arrows) and CCVs (CC). Scale bars: 100 nm. Figure reprinted from Otegui et al. (Otegui et al., 2006) with permission. Anthony L. Contento, and Diane C. Bassham J Cell Sci 2012;125:3511-3518 © 2012. Published by The Company of Biologists Ltd