Single Cell Analysis of Phagocytosis, Phagosome Maturation, Phagolysosomal Leakage, and Cell Death Following Exposure of Macrophages to Silica Particles.

Abstract

Chronic inhalation of silica in various occupational settings results in the development of silicosis, a disease characterized by lung fibrosis. Uptake of silica particles by alveolar macrophages results in cell death and this is one of the contributing factors to the development of silicosis. We have characterized the uncoated or protein-coated (non-opsonized) and Fc receptor-mediated (antibody-opsonized) routes of silica phagocytosis and toxicity. Numerous microscopy techniques and fluorescent probes are outlined in this chapter to carefully measure particle uptake, by macrophages, phagosome maturation, phagosomal reactive oxygen species generation, phagolysosomal leakage, and cell death.