Quantitative Spatiotemporal Analysis of Phagosome Maturation in Live Cells.
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.
Host-Pathogen Interactions in Tuberculosis Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
Laura
L
Schnettger
Host-Pathogen Interactions in Tuberculosis Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK. Maximiliano.Gutierrez@crick.ac.uk.
Maximiliano G
MG
Gutierrez
D000818:Animals; D002470:Cell Survival; D021621:Imaging, Three-Dimensional; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence / Q000379:methods*; D010587:Phagocytosis; D010588:Phagosomes / Q000378:metabolism*; D000067996:RAW 264.7 Cells; D062211:Spatio-Temporal Analysis; D013194:Staining and Labeling
Live cell imaging; Lysosome; Macrophage; Mycobacterium; Phagosome