Analysis of neutrophil membrane traffic during phagocytosis.

Abstract

In this chapter, we describe methods to study membrane traffic during phagosome formation and maturation. Although it is convenient to define events as occurring either before (i.e., during formation) or after (during maturation) the creation of a sealed phagosome, it might not be correct to assume that this is reflected by a sudden change in the membrane traffic events involved. Nevertheless, formation events are studied by approaches different from those of maturation events. Before closure of a phagosome, methods relying on immunochemistry and fluorescent markers are employed. Once phagosomes have formed, these can be isolated and additional methods can be used for their characterization. Here, we describe (1) methods to study membrane traffic during phagosome formation and (2) magnetic purification of bacteria-containing phagosomes for maturation studies.