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Measurement of nonopsonic phagocytic killing by human and mouse phagocytes
Book chapter   Peer reviewed

Measurement of nonopsonic phagocytic killing by human and mouse phagocytes

Richard F Rest and David P Speert
Methods in Enzymology
1994
DOI: https://doi.org/10.1016/0076-6879(94)36010-3
PMID: 7968642
Featured in Collection :   UN Sustainable Development Goals @ Drexel

Abstract

This chapter discusses the methods for the quantitative measurement of phagocytosis and killing of bacteria by human and mouse neutrophils, monocytes, and macrophages. Two common aqueous media are used to isolate neutrophils from blood: Percoll (a suspension of polyvinylpyrrolidone-coated colloidal silica) and a mixture of Ficoll (a nonionic, synthetic sucrose polymer that forms a dense solution) and Hypaque (a dense solution of a polysubstituted carboxylic acid). The chapter presents methods used to investigate phagocytic killing of Neisseria gonorrhoeae and Pseudomonas aeruginosa. The methods work well for almost any bacteria, with a little optimization of the assay. Phagocytic killing of bacteria is measured in a tumbling tube system, where the bacteria and phagocytes are tumbled together, and bacterial viability is measured over time by removing, diluting, and plating aliquots. For phagocytic killing assays, the ratio of bacteria to phagocytes is usually kept rather low, from 0.1 to 2 or 3. Because phagocytes have a limited capacity to internalize and kill their prey, they can become saturated (or satiated).

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19 citations in Web of Science
20 citations in Scopus

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Collaboration types
Domestic collaboration
International collaboration
Web of Science research areas
Biochemical Research Methods
Biochemistry & Molecular Biology
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