Quantitative Spatiotemporal Analysis of Phagosome Maturation in Live Cells

Methods Mol Biol. 2017:1519:169-184. doi: 10.1007/978-1-4939-6581-6_11.

Abstract

Phagocytosis and phagosome maturation are central to the development of the innate and adaptive immune response. Phagosome maturation is a continuous and dynamic process that occurs rapidly. In this chapter, we describe fluorescence-based live cell imaging methods for the quantitative and temporal analysis of phagosome maturation of latex beads and M. tuberculosis as two phagocytic targets. We also describe two simple protocols for monitoring phagosome maturation: the use of the acidotropic probe LysoTracker and analyzing the recruitment of EGFP-tagged host proteins by phagosomes.

Keywords: Live cell imaging; Lysosome; Macrophage; Mycobacterium; Phagosome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Survival
  • Imaging, Three-Dimensional
  • Intracellular Space / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence / methods*
  • Phagocytosis
  • Phagosomes / metabolism*
  • RAW 264.7 Cells
  • Spatio-Temporal Analysis*
  • Staining and Labeling