Conventional transmission electron microscopy

Abstract

Researchers have used transmission electron microscopy (TEM) to make contributions to cell biology for well over 50 years, and TEM continues to be an important technology in our field. We briefly present for the neophyte the components of a TEM-based study, beginning with sample preparation through imaging of the samples. We point out the limitations of TEM and issues to be considered during experimental design. Advanced electron microscopy techniques are listed as well. Finally, we point potential new users of TEM to resources to help launch their project.

FIGURE 1:

A brief flowchart showing the work to be done with different types of sample preparation for conventional electron microscopy (yellow background). The advanced cryo-EM techniques are shown with a blue background. For immuno-EM, the samples can be stained before embedding (pre-embedding staining) or the sections can be stained (post-embedding staining).

FIGURE 2:

Cell structure as visualized by transmission electron microscopy. (A) Actin-myosin cytoskeleton revealed in a cultured cardiomyocyte prepared by conventional chemical fixation. Bar, 1 µm (B) Cytoplasmic organelles in a mouse macrophage prepared by conventional chemical fixation. Bar, 700 nm (C) Golgi membranes in a cultured 3T3 cell prepared by high-pressure freezing and freeze substitution. Bar, 200 nm. (D) Three-dimensional tomographic model of a forming mitotic spindle from budding yeast. Bar, 200 nm.