OPTIMIZATION OF A PHAGOCYTE MICROPLATE CHEMILUMINESCENT ASSAY

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 thi s report we describe the optimization of a lucigenin enhanced phagocyt e chemiluminescent assay using a Titertek Luminoskan. Factors such as cell concentration, serum concentration in the opsonization of the zym osan used and lucigenin concentration were all optimized in our assay. In addition we have found that some of the unique features of the Lum inoskan. continuous microplate agitation during the assay and micropla te temperature control up to 43 degrees C, also significantly enhanced the chemiluminescent response.