Unit

UNIT 14.34 Dynamic Quantitative Assays of Phagosomal Function

  1. Maria Podinovskaia1,
  2. Brian C. VanderVen1,
  3. Robin M. Yates2,
  4. Sarah Glennie3,
  5. Duncan Fullerton4,
  6. Henry C. Mwandumba5,
  7. David G. Russell1

Published Online: 1 OCT 2013

DOI: 10.1002/0471142735.im1434s102

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Podinovskaia, M., VanderVen, B. C., Yates, R. M., Glennie, S., Fullerton, D., Mwandumba, H. C. and Russell, D. G. 2013. Dynamic Quantitative Assays of Phagosomal Function. Current Protocols in Immunology. 102:14.34:14.34.1–14.34.14.

Author Information

  1. 1

    Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York

  2. 2

    Department of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada

  3. 3

    Liverpool School of Tropical Medicine, Liverpool, United Kingdom

  4. 4

    Mid-Cheshire NHS Foundation Trust, Crewe, United Kingdom

  5. 5

    Malawi-Liverpool-Welcome Trust Laboratories, Queen Elizabeth Hospital, Chichiri, Blantyre, Malawi

Publication History

  1. Published Online: 1 OCT 2013

Abstract

Much of the activity of the macrophage as an effector cell is performed within its phagocytic compartment. This ranges from the degradation of tissue debris as part of its homeostatic function to the generation of the superoxide burst as part of its microbicidal response to infection. We have developed a range of real-time readouts of phagosomal function that enable these activities to be rigorously quantified. This unit contains descriptions of several of these assays assessed by different methods of quantitation, including a fluorescence resonance emission transfer (FRET) assay for phagosome/lysosome fusion measured by spectrofluorometry, a fluorogenic assay for the superoxide burst measured by flow cytometry, and a fluorogenic assay for bulk proteolysis measured by confocal microscopy. These assays illustrate both the range of parameters that can be quantified and the flexibility of instrumentation that can be exploited for their quantitation. Curr. Protoc. Immunol. 102:14.34.1-14.34.14. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • macrophage;
  • phagosome;
  • phagocytosis;
  • phagosome maturation