Immunoelectron microscopy

Immunolabelling to localise specific molecules to particular cell organelles can also be applied in electron microscopy. In EM immunolabelling (often known as immunoelectron microscopy, or immuno-EM), the antibody molecules are tagged with electron-dense substances, the most effective being small gold particles, which can be observed as dark dots. Different antibodies can be tagged with particles of different sizes, allowing detection of more than one type of molecule to be performed simultaneously.

Figure 3 shows an example of how immunocytochemistry at the EM level can provide information about the localisation of molecules in an individual cell. The figure shows two cells of a bacterium, Neisseria meningitidis, labelled with an antibody that recognises a particular oligosaccharide (short chain of sugar units). The binding of this primary antibody is visualised using a secondary antibody coupled to gold particles, 15 nm in diameter, each of which can be seen as a very dark (electron-dense) dot.

Figure 3 Electron micrograph showing two cells of the bacterial species Neisseria meningitidis immunolabelled with a primary antibody that recognises an oligosaccharide, followed by a secondary antibody that has been coupled to individual 15 nm gold particles. Note that the cytoplasm has uneven electron density. The DNA in the nucleoid is relatively pale (electron-lucent), while the rest of the cytoplasm is darker (electron-dense) and granular in appearance because it contains many ribosomes.

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The gold particles appear as numerous black dots, all but one of which is on the outer surface of the bacteria. The pale, non-granular cell wall can be distinguished from the darker, granular cytoplasm beneath it. The nucleoid appears as a number of white patches, of a variety of sizes and shapes, within the cytoplasm.

Figure 3 Electron micrograph showing two cells of the bacterial species Neisseria meningitidis immunolabelled with a primary antibody ...