PUBLICATION
Correlative light and electron microscopic detection of GFP-labeled proteins using modular APEX
- Authors
- Ariotti, N., Hall, T.E., Parton, R.G.
- ID
- ZDB-PUB-170523-13
- Date
- 2017
- Source
- Methods in cell biology 140: 105-121 (Chapter)
- Registered Authors
- Hall, Thomas, Parton, Robert G.
- Keywords
- APEX, CLEM, Electron microscopy, GFP-binding peptide, Green fluorescent protein, Light microscopy
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Ascorbate Peroxidases/metabolism*
- Cell Line, Tumor
- Green Fluorescent Proteins/metabolism*
- Microscopy, Electron/methods*
- Subcellular Fractions/metabolism
- Zebrafish
- PubMed
- 28528629 Full text @ Meth. Cell. Biol.
Citation
Ariotti, N., Hall, T.E., Parton, R.G. (2017) Correlative light and electron microscopic detection of GFP-labeled proteins using modular APEX. Methods in cell biology. 140:105-121.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping