An In Vitro Model System to Test Mechano-microbiological Interactions Between Bacteria and Host Cells.
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.
Department of Biology, Brooklyn College of the City University of New York, 307 Ingersoll Hall Extension, 2900 Bedford Avenue, Brooklyn, NY, 11210, USA.|Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA.
Luís Carlos
LC
Santos
Department of Biology, Brooklyn College of the City University of New York, 307 Ingersoll Hall Extension, 2900 Bedford Avenue, Brooklyn, NY, 11210, USA.|Herbert Irving Comprehensive Cancer Center, Columbia University, New York, NY, USA.
Emilia Laura
EL
Munteanu
Department of Biology, Brooklyn College of the City University of New York, 307 Ingersoll Hall Extension, 2900 Bedford Avenue, Brooklyn, NY, 11210, USA. nicolas@mechano-micro-biology.org.|The Graduate Center of the City University of New York, New York, NY, USA. nicolas@mechano-micro-biology.org.
Nicolas
N
Biais
D008841:Actin Cytoskeleton / Q000378:metabolism / Q000382:microbiology; D000199:Actins / Q000378:metabolism; D001696:Biomechanical Phenomena; D002460:Cell Line; D002470:Cell Survival; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology; D054884:Host-Pathogen Interactions; D006801:Humans; D060328:Magnetic Phenomena; D055595:Mechanical Phenomena; D008856:Microscopy, Fluorescence; D009344:Neisseria gonorrhoeae / Q000502:physiology*; D014162:Transfection
D000199:Actins
Cytoskeleton; Magnetic beads; Magnetic tweezers; Neisseria; Type IV pili; f-Actin