The authors describe luminometric analysis of neutrophil respiratory burst in microtitre plates. They analyzed the neutrophil luminescence in small volumes of whole blood and in the so-called buffy coat (plasma with leucocytes after removal of erythrocytes by dextran sedimentation). To produce respiratory burst four types of activators were used: opsonized zymosan, phorbol myristate acetate, N-formyl-Met-Leu-Phe and calcium ionophore A23187. It was revealed that the chemiluminescence response of neutrophils in whole blood and in the buffy coat is very similar in different types of activators, although the chemiluminescence activity of whole blood was always lower as compared with the buffy coat. This was probably due to an increased expression of cell receptors as a result of the separation procedure on the one hand and quenching properties of erythrocytes (haemoglobin) on the other hand. From the assembled results ensues that luminometric analysis in microtitre plates is a reliable protocol for assessment of the neutrophil respiratory burst, whereby assessment in whole blood should be preferred.

Biofyzikální ústav AV CR Brno