Automated image analysis for monitoring oxidative burst in macrophages

Abstract

OBJECTIVE: To evaluate oxidative bursts induced by phorbol myristate acetate in phagocytes at the single-cell level by automated image analysis. STUDY DESIGN: The generation of reactive oxygen species was quantitatively expressed by means of histograms displaying the percentage of cells corresponding to each of the total optical densities measured. RESULTS: Macrophage subpopulations were quantitatively defined. This method allows detailed analysis of the amount of formazan per cell and the sites of deposition of blue precipitate in each cell. CONCLUSION: Image analysis is a reliable quantitative, single-cell assay for studying various cellular characteristics associated with macrophage functions.