Assessing Vacuolar Escape of Listeria Monocytogenes

Methods Mol Biol. 2017:1535:173-195. doi: 10.1007/978-1-4939-6673-8_11.

Abstract

Listeria monocytogenes is a bacterial pathogen which invades and multiplies within non-professional phagocytes. Signaling cascades involved in cellular entry have been extensively analyzed, but the events leading to vacuolar escape remain less clear. In this chapter, we detail a microscopy FRET-based assay which allows quantitatively measuring L. monocytogenes infection and escape from its internalization vacuole, as well as a correlative light/electron microscopy method to investigate the morphological features of the vacuolar compartments containing L. monocytogenes.

Keywords: CCF4; Correlative light/electron microcopy (CLEM); Förster resonance energy transfer (FRET) microscopy; Listeria monocytogenes; Listeriolysin O; Phagocytosis; Phospholipases; Vacuole.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport
  • Fluorescence Resonance Energy Transfer / methods*
  • Listeria monocytogenes / metabolism*
  • Listeria monocytogenes / ultrastructure*
  • Microscopy, Electron / methods*
  • Microscopy, Fluorescence / methods*
  • Vacuoles / metabolism*
  • Vacuoles / ultrastructure