An In Vitro Model System to Test Mechano-microbiological Interactions Between Bacteria and Host Cells

Methods Mol Biol. 2016:1365:195-212. doi: 10.1007/978-1-4939-3124-8_10.

Abstract

The aim of this chapter is to present an innovative technique to visualize changes of the f-actin cytoskeleton in response to locally applied force. We developed an in vitro system that combines micromanipulation of force by magnetic tweezers with simultaneous live cell fluorescence microscopy. We applied pulling forces to magnetic beads coated with the Neisseria gonorrhoeae Type IV pili in the same order of magnitude than the forces generated by live bacteria. We saw quick and robust f-actin accumulation at the sites where pulling forces were applied. Using the magnetic tweezers we were able to mimic the local response of the f-actin cytoskeleton to bacteria-generated forces. In this chapter we describe our magnetic tweezers system and show how to control it in order to study cellular responses to force.

Keywords: Cytoskeleton; Magnetic beads; Magnetic tweezers; Neisseria; Type IV pili; f-Actin.

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Actin Cytoskeleton / microbiology
  • Actins / metabolism
  • Biomechanical Phenomena
  • Cell Line
  • Cell Survival
  • Epithelial Cells / cytology
  • Epithelial Cells / microbiology
  • Host-Pathogen Interactions*
  • Humans
  • Magnetic Phenomena
  • Mechanical Phenomena*
  • Microscopy, Fluorescence
  • Neisseria gonorrhoeae / physiology*
  • Transfection

Substances

  • Actins