Use of a unique chemiluminescence spectrometer in a study of factors influencing granulocyte light emission.

Factors contributing to variability in chemiluminescence (CL) measurements of phagocytizing granulocytes were identified and controlled. Observed CL was decreased by light quenching caused by red blood cells, hemoglobin, phagocytizable particles, and granulocytes. Increases in CL were achieved by opsonization with heat labile and heat stable (antibody) serum factors, reaction temperatures in the range of 37-40 degrees C, and mixing. The effects of controlled temperature variation and mixing were studied with a chemiluminescence spectrometer uniquely designed for such studies. Features of this spectrometer which make it more suitable than the previously employed scintillation spectrometers for the observation of granulocyte and other chemiluminescent systems include; (1) the ability to measure CL immediately upon reaction initiation; (2) simplicity of photomultiplier tube exchange; and (3) built-in optical filter holders for spectral analysis.