title: Optimization of a phagocyte microplate chemiluminescent assay.
abstract: Chemiluminescent assays have been used to quantify phagocytic activity since 1972. In recent years these assays have been adapted to the 96-well microplate format as new luminometers have been developed. In this report we describe the optimization of a lucigenin enhanced phagocyte chemiluminescent assay using a Titertek Luminoskan. Factors such as cell concentration, serum concentration in the opsonization of the zymosan used and lucigenin concentration were all optimized in our assay. In addition we have found that some of the unique features of the Luminoskan, continuous microplate agitation during the assay and microplate temperature control up to 43 degrees C, also significantly enhanced the chemiluminescent response.
authors.affiliation: Medical Countermeasures Section, Defence Research Establishment Suffield, Alberta, Canada.
authors.forename: B
authors.initials: B
authors.lastname: Kournikakis
authors.forename: M
authors.initials: M
authors.lastname: Simpson
mesh_terms: D000166:Acridines; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008163:Luminescent Measurements; D009000:Monocytes / Q000502:physiology*; D009504:Neutrophils / Q000502:physiology*; D010586:Phagocytes / Q000187:drug effects / Q000502:physiology*; D010587:Phagocytosis / Q000187:drug effects / Q000502:physiology; D013696:Temperature; D015054:Zymosan / Q000494:pharmacology
chemical_list: D000166:Acridines; C033472:10,10'-dimethyl-9,9'-biacridinium; D015054:Zymosan