Visualizing non-lytic exocytosis of cryptococcus neoformans from macrophages using digital light microscopy

Sabriya Stukes, Arturo Casadevall

Research output: Contribution to journalArticlepeer-review

Abstract

Many aspects of the infection of macrophages by Cryptococcus neoformans have been extensively studied and well defined. However, one particular interaction that is not clearly understood is non-lytic exocytosis. In this process, yeast cells are released into the extracellular space by a poorly understood mechanism that leaves both the macrophage and Cn viable. Here, we describe how to follow a large number of individually infected macrophages for a 24 hr infection period by time-lapsed microscopy. Infected macrophages are housed in a heating chamber with a CO2 atmosphere attached to a microscope that provides the same conditions as a cell-culture incubator. Live digital microscopy can provide information about the dynamic interactions between a host and pathogen that is not available from static images. Being able to visualize each infected cell can provide clues as to how macrophages handle fungal infections, and vice versa. This technique is a powerful tool in studying the dynamics that are behind a complex phenomenon.

Original languageEnglish (US)
Article numbere52084
JournalJournal of Visualized Experiments
Issue number92
DOIs
StatePublished - Oct 21 2014
Externally publishedYes

Keywords

  • C. neoformans
  • Fungus
  • Host-pathogen interactions
  • Immunology
  • Issue 92
  • Macrophages
  • Non-lytic exocytosis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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