Long-term live cell imaging of cell migration: Effects of pathogenic fungi on human epithelial cell migration

Torsten Wöllert, George M. Langford

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Long-term live cell imaging was used in this study to determine the responses of human epithelial cells to pathogenic biofilms formed by Candida albicans. Epithelial cells of the skin represent the front line of defense against invasive pathogens such as C. albicans but under certain circumstances, especially when the host’s immune system is compromised, the skin barrier is breached. The mechanisms by which the fungal pathogen penetrates the skin and invade the deeper layers are not fully understood. In this study we used keratinocytes grown in culture as an in vitro model system to determine changes in host cell migration and the actin cytoskeleton in response to virulence factors produced by biofilms of pathogenic C. albicans. It is clear that changes in epithelial cell migration are part of the response to virulence factors secreted by biofilms of C. albicans and the actin cytoskeleton is the downstream effector that mediates cell migration. Our goal is to understand the mechanism by which virulence factors hijack the signaling pathways of the actin cytoskeleton to alter cell migration and thereby invade host tissues. To understand the dynamic changes of the actin cytoskeleton during infection, we used long-term live cell imaging to obtain spatial and temporal information of actin filament dynamics and to identify signal transduction pathways that regulate the actin cytoskeleton and its associated proteins. Long-term live cell imaging was achieved using a high resolution, multi-mode epifluorescence microscope equipped with specialized light sources, high-speed cameras with high sensitivity detectors, and specific biocompatible fluorescent markers. In addition to the multi-mode epifluorescence microscope, a spinning disk confocal long-term live cell imaging system (Olympus CV1000) equipped with a stage incubator to create a stable in vitro environment for long-term real-time and time-lapse microscopy was used. Detailed descriptions of these two long-term live cell imaging systems are provided.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Volume1365
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1365
ISSN (Print)10643745

Keywords

  • Actin cytoskeleton
  • Biofilms
  • Candida albicans
  • Cell migration
  • Environmental control chambers
  • Keratinocytes
  • Live cell imaging
  • Real-time and time-lapse observations

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Wöllert, T., & Langford, G. M. (2016). Long-term live cell imaging of cell migration: Effects of pathogenic fungi on human epithelial cell migration. In Methods in Molecular Biology (Vol. 1365). (Methods in Molecular Biology; Vol. 1365). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3124-8_1