Single-molecule imaging in living cells can provide unique information about biological processes. Bacteria offer some particular challenges for single-molecule imaging due to their small size, only slightly larger than the diffraction limit of visible light. Here, we describe how reliable and reproducible single-molecule data can be obtained for a transmembrane protein in the Gram-negative bacterium Escherichia coli by using live-cell fluorescence microscopy. Fluorescent labeling of a protein by genetic fusion, cell culturing, sample preparation, imaging, and data analysis are discussed.
CITATION STYLE
Varadarajan, A., Oswald, F., & Bollen, Y. J. M. (2018). Single-molecule imaging of Escherichia coli transmembrane proteins. In Methods in Molecular Biology (Vol. 1665, pp. 135–144). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7271-5_7
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