A Rapid, Specific Assay for Superoxide Release from Phagocytes in Small Volumes of Whole Blood

Michael F. Tosi, Akbar Hamedani
1992 American Journal of Clinical Pathology  
A rapid, specific assay was developed to measure the release of superoxide anion from stimulated phagocytic cells in small volumes of whole blood. The assay is based on the chemiluminescence that results when lucigenin (bis-N-methyl acridinium nitrate) is reduced by superoxide. Heparinized whole blood, at volumes from 0.2 mL to a single drop from a 20-gauge needle, was mixed with lucigenin and either a soluble or particulate stimulus (phorbol myristate acetate or opsonized zymosan particles,
more » ... pectively) in a standard volume of 1 mL. The chemiluminescence was measured at 3-minute intervals for a 30-minute period in a luminometer capable of automated operation. Characteristic plots of chemiluminescence versus time were obtained. This assay is rapid and simple, obviates the need to isolate leukocytes from whole blood, is specific for superoxide, can be
doi:10.1093/ajcp/97.4.566 pmid:1313205 fatcat:vsj2d2dwkrhjfa6uyzotqbn67e