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, 140, 105-121

Correlative Light and Electron Microscopic Detection of GFP-labeled Proteins Using Modular APEX

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Correlative Light and Electron Microscopic Detection of GFP-labeled Proteins Using Modular APEX

Nicholas Ariotti et al. Methods Cell Biol.

Abstract

The use of green fluorescent protein (GFP) and related proteins has revolutionized light microscopy. Here we describe a rapid and simple method to localize GFP-tagged proteins in cells and in tissues by electron microscopy (EM) using a modular approach involving a small GFP-binding peptide (GBP) fused to the ascorbate peroxidase-derived APEX2 tag. We provide a method for visualizing GFP-tagged proteins by light and EM in cultured cells and in the zebrafish using modular APEX-GBP. Furthermore, we describe in detail the benefits of this technique over many of the currently available correlative light and electron microscopy approaches and demonstrate APEX-GBP is readily applicable to modern three-dimensional techniques.

Keywords: APEX; CLEM; Electron microscopy; GFP-binding peptide; Green fluorescent protein; Light microscopy.

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