Single-molecule imaging of Escherichia coli transmembrane proteins

A. Varadajan, F. Oswald, Y.J.M. Bollen*

*Corresponding author for this work

Research output: Chapter in Book / Report / Conference proceedingChapterAcademicpeer-review

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
EditorsErwin Peterman
PublisherHumana Press Inc
Pages135-144
Number of pages10
Volume1665
ISBN (Electronic)9781493972715
ISBN (Print)9781493972708
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1665
ISSN (Print)1064-3745

Keywords

  • Bacteria
  • Diffusion
  • Escherichia coli
  • Fluorescence microscopy
  • Sample preparation
  • Single-molecule tracking
  • Transmembrane protein

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  • Cite this

    Varadajan, A., Oswald, F., & Bollen, Y. J. M. (2018). Single-molecule imaging of Escherichia coli transmembrane proteins. In E. Peterman (Ed.), Methods in Molecular Biology (Vol. 1665, pp. 135-144). (Methods in Molecular Biology; Vol. 1665). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7271-5_7