Luminometric Analysis of the Neutrophils Respiratory Burst inMicrotitre Plates

Overview

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-calledbuffy coat (plasma with leucocytes after removal of erythrocytes by dextran sedimentation).To produce respiratory burst four types of activators were used: opsonized zymosan, phorbolmyristate acetate, N-formyl-Met-Leu-Phe and calcium ionophore A23187. It was revealed that thechemiluminescence response of neutrophils in whole blood and in the buffy coat is very similar indifferent types of activators, although the chemiluminescence activity of whole blood was alwayslower as compared with the buffy coat. This was probably due to an increased expression of cellreceptors as a result of the separation procedure on the one hand and quenching properties oferythrocytes (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 respiratoryburst, whereby assessment in whole blood should be preferred.

Key words:
chemiluminescence – neutrophils – respiratory burst.