Use of BMG LABTECH microplate reader to monitor the production of free radicals by leukocytes in diluted blood

In the course of an immune response, reactive oxygen species (ROS) are produced and released during a respiratory burst in order to damage pathogens. ROS can be measured using Pholasin^®, the photoprotein of the marine rockboring bioluminescent mollusk. The protein glows upon activation by free radicals and is used in the ABEL^® Antioxidant kit.

The kit was used in combination with a luminescence-capable BMG LABTECH plate reader to determine stimulation of leukocytes in whole blood. Blood samples were stimulated with either the tetrapeptide fMLP or with phorbol 12-myristate 13-acetate (PMA). Upon addition of the stimulant by on-board reagent injectors, ROS response was monitored. Differences in ROS-production upon fMLP and PMA were detected. The monitoring of leukocytes over hours was enabled by the temperature control integrated in the reader.